CN101685089A - Method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs - Google Patents
Method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs Download PDFInfo
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Abstract
The invention discloses a method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs. In the method, 70-percent methanol is used as a sample ultrasonic extraction solvent, a C18 chromatographic column is adopted, a system of acetonitrile-tetrahydrofuran-formic acid-water is used as a flowing phase, an electrospray ionization(ESI) ion source is adopted, quick separation, identification and measurement of pseudoginsenoside F11 and ginsenoside Rf which are characteristics components of the American ginsengs and ginsengs respectively are performed within 2 minutes for the first time under a condition of a negative ion mode second order mass spectrometry analysis, and whether the preparation of the American ginsengs is mixed with the ginsengs is judged and the content of crude drugs mixed with the ginsengs is worked out. The method has the advantages of strong specificity, synchronous qualitative and quantitative analysis, easy and quick pre-treatment, quick detection of the contents of the American ginsengs and the ginsengs within 2 minutes, high sensitivity of detection and measurement and good repeatability, andthe suitability for quick quality screening of the American ginsengs, the ginsengs and products of the American ginsengs and the ginsengs.
Description
Technical field
The invention belongs to Chinese crude drug detection technique field, relate to the quality testing of Chinese medicine medicine, health products, particularly use Ultra Performance Liquid Chromatography-level Four bar-flight time mass spectrum coupling to American Ginseng and genseng and preparation is differentiated fast and the method for quantitative measurement.
Background technology
American Ginseng (Panax quinque folium L.) and genseng (Panax ginseng C.A.Mey) belong to Araliaceae together.The American Ginseng main product has boosting qi and nourishing yin in North America, effects such as clearing heat and promoting fluid.China also introduces a fine variety the Northeast, is commonly called as to be kind of an American ginseng.Panax per nnial herb, main product have and reinforce vital energy in China the Northeast, and multiple arteries and veins takes off admittedly, reinforces the spleen to benefit the lung, and is promoting production of body fluid and inducing sedation of the mind, effects such as happy intelligence development.The two principal ingredient is the ginsenoside compounds, but drug effect and price differ bigger.Traditional Chinese medical theory is thought, sweet, the little hardship of American Ginseng flavor is cool in nature, sweet, the little hardship of genseng flavor, and property is flat.American Ginseng has raising immunity, and various goods can directly be taken or make to constitutional effect, as products such as capsule, lozenge, tea bag, electuary and oral liquids, can be used as daily health products, the huge market demand.Just there are nearly ten thousand families in the manufacturing enterprise of the American ginseng buccal tablet of whole nation production at present.But because import ginseng cost height, and output is limited, far can not satisfy the production demand, domestic in addition genseng relative low price, and the composition mouthfeel is close, is difficult for difference, cause that adulterate in American Ginseng market, the fake and inferior commodities ubiquity, the grievous injury consumer's interests.And pharmacopeia act.std all can not effectively be differentiated the American Ginseng gen-seng both at home and abroad.Demand setting up American Ginseng or genseng and preparation discriminating assay method thereof fast and effectively urgently.
Existing achievement in research shows that the kind of American Ginseng and genseng ingredient is close, but its characteristic component is respectively arranged.American Ginseng contain the characteristic component pseudo-ginsenoside F 11, only contain 1,000,000 in the genseng/; And contained characteristic component ginsenoside Rf in the genseng does not exist in the American Ginseng.But pseudo-ginsenoside F 11 and ginsenoside Rf belong to isomers, and polarity is closely similar, and molecular weight is identical, generally are difficult to separate, and cause distinguishing difficulty.Existing analytical approach sample preparation complexity, detection time is generally more than 60min, and separating effect is not good, is unfavorable for detection by quantitative and rapid screening.
" Chinese pharmacopoeia has adopted the thin layer differential method to the discriminating of American ginseng medicine to existing 2005 editions, using American Ginseng control medicinal material, pseudo-ginsenoside F 11 reference substance, ginsenoside Rb1's reference substance, ginsenoside Re's reference substance, reaching ginsenoside Rg1's reference substance is contrast, wherein pseudo-ginsenoside F 11 is the characteristic component that American Ginseng is different from genseng, but contained ginsenoside Rf is an isomers in this composition and the genseng, be difficult to distinguish with thin-layer chromatography, so list is not enough to differentiate American Ginseng with this method.Also only adopt high performance liquid chromatography under detection wavelength 203nm, to measure under the assay item in the standard, do not relate to the characteristic component of American Ginseng yet ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1.And preparation method's complexity of need testing solution needs with water saturated normal butyl alcohol reflux 1.5 hours, put cold, mend heavy after, evaporate to dryness, with the redissolution of 50% methyl alcohol, constant volume is diluted in transfer more again.Separating through the C18 post, is moving phase with acetonitrile-0.1% phosphoric acid solution, and behind the gradient elution, the 203nm wavelength is measured absorbance log down, and single Liquid Detection time just needs more than the 100min, the method complicated and time consumption, and specificity is not strong, still is difficult to American Ginseng is differentiated detection.Quick method of separating and assaying to pseudo-ginsenoside F 11 and ginsenoside Rf does not appear in the newspapers as yet at present.
Ultra Performance Liquid Chromatography is compared common liquid chromatography, and it is fast to have an analysis speed, and separating power is strong, good reproducibility, and characteristics such as sample size is little are fit to and difficult separation component are separated fast.That the liquid chromatograph mass spectrography technology has realized simultaneously is strong to boiling point height, polarity, heat-labile compound liquid phase separation and qualitative and quantitative detection, highly sensitive, detection limit is low, compound structure information and content information can be provided, reach the purpose of sample being carried out simultaneously qualitative, quantitative, strengthen selectivity, shortened analysis time.The combination of level Four bar-flight time mass spectrum more can the lock onto target ion, carries out the parsing of second order ms, and has high resolving power, and the molecular weight of pinpoint accuracy is provided.At present, liquid chromatography-mass spectrography-mass spectrometry has developed into a kind of important analytical technology, and the application in many fields such as biological food, environmental chemistry, biological chemistries launches gradually.The key of its analysis condition is the selection of moving phase and the optimization of mass spectrum condition, as collision energy, nitrogen flow rate and source temperature etc.
Summary of the invention
The objective of the invention is to set up a kind of detection method of differentiating fast and effectively, promptly the quality true and false of American Ginseng preparation is carried out fast assessing, the method that a kind of sample treatment is easy, detect pseudo-ginsenoside F 11 and ginsenoside Rf in American Ginseng and the genseng when shortening analysis time and low detectability is provided, and according to existing The effects data, can calculate the content that genseng mixes in the American Ginseng preparation, be a kind of method that the American Ginseng product is carried out the rapid evaluation investigation, be applicable to that simultaneously all various formulation products that contain American Ginseng and genseng detect.For finishing above-mentioned purpose, the present invention discloses following technical scheme:
A kind of method of utilizing the Ultra Performance Liquid Chromatography-level Four bar-flight time mass spectrum coupling fast detecting American Ginseng genseng and the quality of the pharmaceutical preparations true and false thereof mainly comprises sample preparation, liquid phase separation, three aspects of Mass Spectrometer Method, and step is as follows down for it:
(1) preparation of need testing solution: get this product American ginseng buccal tablet powder 1g, cross 40 mesh sieves, accurately claim surely, add 70-95% methyl alcohol 50ml, ultrasonic Extraction 20min takes out, and puts coldly, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, and gets subsequent filtrate, and is standby;
(2) preparation of reference substance solution: precision takes by weighing pseudo-ginsenoside F
11Reference substance, ginsenoside Rf's reference substance are an amount of, add methyl alcohol and make every 1mL respectively and contain pseudo-ginsenoside F
112.028ng, the solution of ginsenoside Rf 0.684ng, standby;
(3) Ultra Performance Liquid Chromatography-mass spectrometry combination method qualitative identification:
Chromatographic condition: adopt Acquity UPLC BEH C18 chromatographic column (2.1 * 100mm, 1.7 μ m); Liquid-phase chromatographic column, with mobile phase A acetonitrile-tetrahydrofuran 10: 1, Mobile phase B 0.2% formic acid water carried out isocratic elution, flow velocity 0.4mL/min, 30 ℃ of column temperatures; Need testing solution input 5 μ l.
The wherein preferred mobile phase A acetonitrile-tetrahydrofuran (10: 1) that adopts: the volume ratio of B 0.2% formic acid water=30: 70.
Mass spectrum condition: adopt the electron spray ionisation source, negative ion electrospray is from pattern, kapillary ionization voltage 2.4kV, sampling taper hole voltage 15V, 120 ℃ of ion source temperatures, the desolventizing temperature: 350 ℃, desolventizing nitrogen flow rate 600L/hr, taper hole blowback nitrogen 50L/hr, level Four bar sweep limit m/z 250～1000Da, acquisition time 0～8min, sweep time 0.3s, interval time 0.02s, select ion m/z 835Da as object ion, collision voltage 45v, TOF ion flight mode adopts V model, use LEK m/z 554.2615, object ion is carried out the accurate mass locking as external standard; Judge whether contain pseudo-ginsenoside F according to gained second order ms figure
11Secondary fragment ion peak group with the ginsenoside Rf;
(4) pseudo-ginsenoside F
11With ginsenoside Rf's quantitative measurement:
The same step of liquid phase process (3) is described, and reference substance solution and need testing solution be sample introduction 5 μ l respectively, and the same step of mass spectrometry method (3) is described, collision voltage 15v, and integration in reference substance mass spectrum chromatogram and sample mass spectrum chromatogram is measured pseudo-ginsenoside F
11The peak area of mass spectrum chromatographic peak and ginsenoside Rf's mass spectrum chromatographic peak;
(5) pseudo-ginsenoside F in the calculation sample
11With ginsenoside Rf's content, and according to pseudo-ginsenoside F in known American Ginseng and the genseng
11With ginsenoside Rf's content data, measure American Ginseng and ginseng crude drug's content in the test sample.
Quick discriminating detection method of the present invention is preferably got this product American ginseng buccal tablet powder 1g, crosses 40 mesh sieves, and accurate the title decides, add 70% methyl alcohol 50ml, ultrasonic Extraction 20min takes out, and puts cold, add 70% methyl alcohol and supply the weight that subtracts mistake, shake up, filter, get subsequent filtrate.
Quick discriminating detection method of the present invention, wherein pseudo-ginsenoside F
11Relative retention time at 1.5min, and ginsenoside Rf's relative retention time is at 1.7min, the two reaches baseline separation.
Quick discriminating detection method of the present invention, at collision voltage is under the 4.5v, extract the second order ms figure of quasi-molecular ions m/z 835, as the characteristic component ginsenoside Rf's of genseng feature fragment peak group m/z799, m/z637, m/z475 appear, fragment peak group m/z799, m/z653, m/z491 are the characteristic component pseudo-ginsenoside F of American Ginseng
11Fragmention.
Quick discriminating detection method of the present invention, pseudo-ginsenoside F wherein
11With ginsenoside Rf's detection method can the identical or close any formulation that contains American Ginseng, genseng of Detection and Extraction method.For example tablet, capsule or granule.
Quick discriminating detection method of the present invention, preferred 70% methyl alcohol adopts ultrasonic extracting method, ultrasonic 20min for extracting solvent.Good to the target component extraction effect, and method is simple to operation, experimental result, and the recovery of this method, repeatability and extraction efficiency all reach requirement.
One of gordian technique of the present invention is these two kinds of isomerss of pseudo-ginsenoside F 11 and ginsenoside Rf were realized separating fast in 2 minutes, adopt mobile phase A-B (30: 70), wherein mobile phase A is acetonitrile-tetrahydrofuran (10: 1), Mobile phase B is 0.2% formic acid water, wherein add tetrahydrofuran and can strengthen the retention of different component in post in various degree, measure that separating effect is not obvious at least, amount can not further increase degree of separation at most, and make each component retention time prolong, increase detection time.The rarely seen moving phase in Liquid Detection of tetrahydrofuran is selected, and effect is remarkable in the present invention.
Adopt level Four bar-flight time mass spectrum that quasi-molecular ions m/z835 is carried out second order ms and detect, the selection of collision energy is most important.Energy is low excessively, and quasi-molecular ions m/z835 is not enough to fragmentation, and it is low to form the fragmention abundance; Energy is too high, and the characteristic ion peak group m/z799 of pseudo-ginsenoside F 11, m/z653, m/z491 and ginsenoside Rf's characteristic ion peak group m/z799, m/z637, m/z475 are also broken thereupon, can't differentiate.Be applicable to that collision voltage of the present invention is 45v, selection m/z835 is an object ion, the fragmention peak area maximum that each stack features quasi-molecular ions can detect and form this moment.
The present invention shows through methodological study, with 70% methyl alcohol ultrasonic Extraction American Ginseng preparation, adopting Ultra Performance Liquid Chromatography-level Four bar-flight time mass spectrum method for combined use to differentiate detects, wherein pseudo-ginsenoside F 11 and ginsenoside Rf and detection limit reach 0.08ng and 0.05ng respectively, quantitative limit is respectively 0.243ng and 0.164ng, the recovery is respectively 97.24% (RSD=4.6, n=6) and 93.75% (RSD%=4.4, n=6), the range of linearity is respectively 0.164～16.4ng (linear equation y=8.2405x+2.1062, r=0.9989) and 0.243～24.3ng (linear equation y=8.7416x+1.8629, r=0.9997); The samples of Ginseng of mixing in the American Ginseng of self-control different proportion (adding ginseng crude drug's ratio 1%, 5%, 10%, 30%, 50%, 75%, 100%), measured deviation is 0.09～3.85%.
In order to pass through detection by quantitative result to pseudo-ginsenoside F 11 in the sample and two kinds of compositions of ginsenoside Rf, more accurately infer the wherein content of American Ginseng and genseng crude drug, thereby draw the ratio of doping genseng, collected 16 batches of the American ginseng medicines in the different places of production on the market and ginseng crude drug 11 batches among the present invention, respectively pseudo-ginsenoside F 11 and ginsenoside Rf's content is wherein measured, obtained the content mean value of ginsenoside Rf in the content mean value of pseudo-ginsenoside F 11 in the American ginseng medicine and the genseng.See Table 2,3.
Each batch of table 2. American Ginseng sample determination result
| The American Ginseng sample number into spectrum | The American Ginseng sample place of production | Pseudo-ginsenoside F 11 content (μ g/g) |
| ??xys0001 | Canada | ??1021.8 |
| ??xys0002 | Wendeng City | ??1062.4 |
| ??xys0003 | Beijing | ??827.28 |
| ??xys0101 | Tonghua, Jilin | ??727.51 |
| ??xys0201 | The Huadian, Jilin | ??689.71 |
| ??xys0301 | Laiyang, Shandong | ??1313.6 |
| ??xys0401 | Huairou, Beijing | ??1096.6 |
| ??xys0601 | The Qingdao | ??1171.1 |
| ??xys0701 | Canada, the Ontario | ??1211.7 |
| ??xys1301 | Beautiful Country, the Wisconsin State | ??987.72 |
| ??Xys1401 | Beautiful Country, the Wisconsin State | ??978.62 |
| ??xys1901 | Canada, inferior Poetry economizes | ??862.27 |
| ??Lxys0101 | China | ??1100.1 |
| ??Lxys0201 | Canada, the Ontario | ??984.76 |
| ??Lxys0301 | Beautiful Country, the Wisconsin State | ??1097.7 |
| ??Lxys0401 | Canada, inferior Poetry economizes | ??787.06 |
| Mean value | ??995.00 |
Each batch of table 3. samples of Ginseng measurement result
| The samples of Ginseng numbering | Samples of Ginseng content | ??Rf(μg/g) |
| ??Rs001 | Tongrentang | ??631.93 |
| ??Rs002 | The Jingyu, Jilin | ??267.89 |
| ??Rs003 | Liaoning Huan Ren Mali pasture | ??529.14 |
| ??Rs004 | The Fusong horse is good in Jilin | ??266.51 |
| ??Rs005 | Ceng Fuwang | ??611.65 |
| ??Rs006 | Ji'an, Jilin | ??520.09 |
| ??Rs007 | Yu Rensheng | ??416.32 |
| ??Rs008 | The Antu, Jilin | ??563.20 |
| ??Rs009 | The Jilin Fusong | ??485.64 |
| ??Rs010 | Dongfanghong | ??770.44 |
| ??Rs011 | Genseng (having by oneself) | ??475.62 |
| Mean value | ??503.49 |
The present invention compares the good effect that is had with existing detection technique and is:
(1) realize first American Ginseng and genseng separately characteristic component pseudo-ginsenoside F 11 and the quick of ginsenoside Rf separate, can finish detection in the 2min.
(2) adopt second order ms, find the fragment ion peak of feature separately of two kinds of compositions, can differentiate evaluation to component simultaneously.
(3) extracting method is simple, method strong operability, recovery height, good reproducibility.
(4) through experimental verification, this method accuracy height has accumulated reliable experimental, can infer preparation Chinese crude drug addition and additional proportion by the assay to single component.
Description of drawings
Fig. 1 pseudo-ginsenoside F 11 reference substance and ginsenoside Rf's reference substance TIC figure, retention time is respectively at 1.5min and 1.7min (TOF MSMS ES-TIC);
The TIC figure (TOF MSMS ES-TIC) of Fig. 2 American ginseng tea sample;
The second order ms figure of Fig. 3 pseudo-ginsenoside F 11 (TOF MSMS 835.00ES.228);
Fig. 4 ginsenoside Rf's second order ms figure (TOF MSMS 835.00ES.228).
Embodiment
Below in conjunction with preferred embodiment, to being described in detail as follows according to embodiment provided by the invention; Simultaneously for simple and purpose clearly, hereinafter appropriate omission the description of known technology, in order to avoid those unnecessary details influences are to the description of the technical program.
Embodiment 1
1, material
1.1 instrument and material
U.S. Waters ACQUITY Ultra Performance LC Ultra Performance Liquid Chromatography instrument; Waters Q-TOF Premier high-resolution quadrupole rod and flight time tandem mass spectrometer; Be equipped with electric spray ion source (ESI) and Masslynx 4.1 data acquisition softwares.
1.2 medicine and reagent
03091112), pseudo-ginsenoside F 11 reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 841-9903) ginsenoside Rf (Extrasynthese company, lot number:; Acetonitrile (chromatographically pure, U.S. Fisher company), formic acid (chromatographically pure, U.S. Fisher company), tetrahydrofuran (chromatographically pure, U.S. Fisher company); Ultrapure water (0.22 μ m filter membrane is crossed by milli Q ultrapure water system).The propylhomoserin enkephalins is bought the Sigma company in the U.S..
2. method
Get American ginseng buccal tablet powder (cross 40 mesh sieves) 1g, accurately claim surely, add 70% methyl alcohol 50ml, ultrasonic Extraction 20min takes out, and puts coldly, adds 70% methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, and gets subsequent filtrate, promptly.
The preparation of reference substance solution: precision takes by weighing the pseudo-ginsenoside F 11 reference substance, ginsenoside Rf's reference substance is an amount of, adds methyl alcohol and makes the solution that every 1mL contains pseudo-ginsenoside F 11 2.028ng, ginsenoside Rf 0.684ng respectively, promptly; Qualitative identification:
Liquid-phase condition, chromatographic column: Acquity UPLC BEH C
18Chromatographic column (2.1 * 100mm, 1.7 μ m); Moving phase table 1; Flow velocity 0.4ml/min; 30 ℃ of column temperatures; Need testing solution sample size 5 μ l, sample chamber temperature: 4 ℃; Room temperature: 24 ℃.
Table 1 moving phase
Q-TOF MS condition: adopt the electron spray ionisation source, negative ion electrospray is from pattern, kapillary ionization voltage 2.4kV, and sampling taper hole voltage 15V, 120 ℃ of ion source temperatures, the desolventizing temperature: 350 ℃, desolventizing nitrogen flow rate 600L/hr, taper hole blowback nitrogen 50L/hr.Level Four bar sweep limit m/z 250～1000Da, acquisition time: 5min, sweep time: 0.3s, interval time 0.02s, select ion m/z 835Da as object ion, collision voltage 45v, TOF ion flight mode adopts V model, uses LEK (m/z 554.2615) as external standard object ion to be carried out the accurate mass locking.Judge the secondary fragment ion peak group that whether contains the ginsenoside Rf according to gained second order ms figure.
Quantitative measurement: reference substance solution and need testing solution be sample introduction 5 μ l respectively, detection method is with above-mentioned qualitative identification, collision voltage changes 15v into, integration in reference substance mass spectrum chromatogram and sample mass spectrum chromatogram, the peak area of mensuration pseudo-ginsenoside F 11 mass spectrum chromatographic peak and ginsenoside Rf's mass spectrum chromatographic peak.Pseudo-ginsenoside F 11 and ginsenoside Rf's content in the calculation sample, and, measure American Ginseng and ginseng crude drug's content and ratio in the test sample according to the content data of pseudo-ginsenoside F 11 and ginsenoside Rf in known American Ginseng and the genseng.
2.1 the range of linearity
Make the ginsenoside Rf and the pseudo-ginsenoside F 11 reference substance solution of variable concentrations respectively, enter the LC-MS instrument respectively, measure by above-mentioned quantitative measurement condition, in the mass spectrum chromatogram that forms, extract m/z835 ion flow graph, peak area is carried out integration, (μ g) is horizontal ordinate x with sample size, is ordinate y with the peak area, carries out regression treatment.Ginsenoside Rf's regression equation y=8.7416x+1.8629, r=0.9997; Pseudo-ginsenoside F 11 regression equation y=8.2405x+2.1062, r=0.9989.The result shows, ginsenoside Rf and pseudo-ginsenoside F 11 are good linear relationship at 0.164～16.4ng and 0.243～24.3ng sample size and peak area respectively.
2.2 precision
The sample of getting same lot number is according to need testing solution preparation method preparation, 6 parts of need testing solutions of parallel preparation are pressed the said determination method and are measured, and calculate ginsenoside Rf and pseudo-ginsenoside F 11 content, RSD (%) is respectively 4.00 and 1.08 as a result, shows this method good reproducibility.
2.3 the recovery
Get 6 parts in the sample (ginsenoside Rf 10.01 μ g/g, pseudo-ginsenoside F 11 50.45 μ g/g) of known content, every part of about 0.5g, to 100ml tool plug conical flask, accurate respectively 70% methanol solution (containing ginsenoside Rf's reference substance 0.1026 μ g/ml and the pseudo-ginsenoside F 11 reference substance 0.507 μ g/ml) 50ml that adds, the accurate 70% methyl alcohol 50ml that adds claims to decide weight, sonicated 20min, put coldly, supply the weight that subtracts mistake, filter with 70% methyl alcohol, get subsequent filtrate, sample introduction is measured.Calculate ginsenoside Rf and pseudo-ginsenoside F 11 average recovery rate and be respectively 97.24% and 93.75%, RSD (%) is respectively 4.39 and 4.61 (n=6).
3. result
Measured the content of ginsenoside Rf and pseudo-ginsenoside F 11 in 2 kinds of American Ginseng preparations that commercially available A produces house, standard content in contained ginsenoside Rf and pseudo-ginsenoside F 11 in ginseng crude drug and the American ginseng medicine, calculate the ratio of contained ginseng crude drug and American ginseng medicine in the American Ginseng preparation, testing result (seeing Table 4)
The commercially available A of table 4. producer American Ginseng preparation measurement result
Embodiment 2
Method is with embodiment 1.
Measured the content of ginsenoside Rf and pseudo-ginsenoside F 11 in 2 kinds of American Ginseng preparations that commercially available B produces house, standard content in contained ginsenoside Rf and pseudo-ginsenoside F 11 in ginseng crude drug and the American ginseng medicine, calculate the ratio of contained ginseng crude drug and American ginseng medicine in the American Ginseng preparation, testing result (seeing Table 5)
The commercially available B of table 5. producer American Ginseng preparation measurement result
After the preferred embodiment that describes in detail, being familiar with this technology personage can be well understood to, can carry out various variations and modification not breaking away under above-mentioned claim and the spirit, all foundations technical spirit of the present invention all belongs to the scope of technical solution of the present invention to any simple modification, equivalent variations and modification that above embodiment did.And the embodiment that the present invention also is not subject in the instructions to be given an actual example.
Claims (7)
1, a kind of method of utilizing the Ultra Performance Liquid Chromatography-level Four bar-flight time mass spectrum coupling fast detecting American Ginseng genseng and the quality of the pharmaceutical preparations true and false thereof is characterized in that this method may further comprise the steps successively:
(1) preparation of need testing solution: get this product American ginseng buccal tablet powder 1g, accurate claim surely, ultrasonic Extraction 20-30min takes out, and puts coldly, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, and gets subsequent filtrate, and is standby;
(2) preparation of reference substance solution: precision takes by weighing pseudo-ginsenoside F
11Reference substance, ginsenoside Rf's reference substance are an amount of, add methyl alcohol and make every 1mL respectively and contain pseudo-ginsenoside F
112.028ng, the solution of ginsenoside Rf 0.684ng, standby;
(3) Ultra Performance Liquid Chromatography-mass spectrometry combination method qualitative identification:
Chromatographic condition: adopt C
18Liquid-phase chromatographic column; With mobile phase A acetonitrile-tetrahydrofuran 10: 1, B 0.2% formic acid water carried out isocratic elution, flow velocity 0.4mL/min, 30 ℃ of column temperatures; Need testing solution input 5 μ l.
Mass spectrum condition: adopt the electron spray ionisation source, negative ion electrospray is from pattern, kapillary ionization voltage 2.4kV, sampling taper hole voltage 15V, 120 ℃ of ion source temperatures, the desolventizing temperature: 350 ℃, desolventizing nitrogen flow rate 600L/hr, taper hole blowback nitrogen 50L/hr, level Four bar sweep limit m/z 250～1000Da, acquisition time 0～8min, sweep time 0.3s, interval time 0.02s, select ion m/z 835Da as object ion, collision voltage 45v, TOF ion flight mode adopts V model, use LEK m/z 554.2615, object ion is carried out the accurate mass locking as external standard; Judge whether contain pseudo-ginsenoside F according to gained second order ms figure
11Secondary fragment ion peak group with the ginsenoside Rf;
(4) pseudo-ginsenoside F
11With ginsenoside Rf's quantitative measurement:
The same step of liquid phase process (3) is described, and reference substance solution and need testing solution be sample introduction 5 μ l respectively, and the same step of mass spectrometry method (3) is described, collision voltage 15v, and integration in reference substance mass spectrum chromatogram and sample mass spectrum chromatogram is measured pseudo-ginsenoside F
11The peak area of mass spectrum chromatographic peak and ginsenoside Rf's mass spectrum chromatographic peak;
(5) pseudo-ginsenoside F in the calculation sample
11With ginsenoside Rf's content, and according to pseudo-ginsenoside F in known American Ginseng and the genseng
11With ginsenoside Rf's content data, measure American Ginseng and ginseng crude drug's content in the test sample.
2, method for quick according to claim 1 is characterized in that getting this product American ginseng buccal tablet powder 1g, crosses 40 mesh sieves, accurate claim fixed, add 70% methyl alcohol 50ml, ultrasonic Extraction 20min takes out, and puts cold, add 70% methyl alcohol and supply the weight that subtracts mistake, shake up, filter, get subsequent filtrate.
3, method for quick as claimed in claim 1, the volume ratio A of its step (3) moving phase: B=30: 70.
4, method for quick as claimed in claim 1, wherein pseudo-ginsenoside F
11Relative retention time at 1.5min, and ginsenoside Rf's relative retention time is at 1.7min, the two reaches baseline separation.
5, method for quick as claimed in claim 1 is under the 4.5v at collision voltage, extracts the second order ms figure of quasi-molecular ions m/z 835; Wherein constituent of ginseng ginsenoside Rf's feature fragment ion peak group is m/z799, m/z637, m/z475; The fragment ion peak group of American Ginseng composition pseudo-ginsenoside F 11 is m/z799, m/z653, m/z491.
6, as claims 1～5 each described method for quick, pseudo-ginsenoside F wherein
11With ginsenoside Rf's detection method can the identical or close any formulation that contains American Ginseng, genseng of Detection and Extraction method.
7, as claims 6 described method for quick, formulation wherein comprises tablet, capsule or granule.
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|---|---|---|---|
| CN 200910069378 CN101685089B (en) | 2009-06-23 | 2009-06-23 | Method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs |
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|---|---|---|---|
| CN 200910069378 CN101685089B (en) | 2009-06-23 | 2009-06-23 | Method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs |
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| CN101685089A true CN101685089A (en) | 2010-03-31 |
| CN101685089B CN101685089B (en) | 2013-01-02 |
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| CN 200910069378 Expired - Fee Related CN101685089B (en) | 2009-06-23 | 2009-06-23 | Method for quickly quality-detecting and identifying American ginsengs, ginsengs and preparations of American ginsengs and ginsengs |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103592376A (en) * | 2013-07-02 | 2014-02-19 | 牡丹江友搏药业股份有限公司 | Novel quality control method of Shuxuetong injection |
| CN105301162A (en) * | 2015-11-10 | 2016-02-03 | 天津市尖峰天然产物研究开发有限公司 | Method for measuring pseudo-ginsenoside F11 in American ginseng through pre-column derivatization high performance liquid chromatography |
| CN106645482A (en) * | 2016-12-26 | 2017-05-10 | 河北省药品检验研究院 | Method for determining content of four ginsenoside components in 27-flavor Dingkun pill |
| CN107064329A (en) * | 2016-12-26 | 2017-08-18 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of Xueshuan xinmaining Tablet |
| CN107064328A (en) * | 2016-12-26 | 2017-08-18 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of ginseng spleen-strengthening bolus |
| CN110568096A (en) * | 2019-09-09 | 2019-12-13 | 哈尔滨工业大学（威海） | rapid detection method for multiple saponins of American ginseng |
| CN114113444A (en) * | 2021-11-24 | 2022-03-01 | 天津中医药大学 | Method for identifying traditional Chinese medicine derived from Panax plant or different parts thereof and application thereof |
| US11402325B2 (en) | 2021-01-15 | 2022-08-02 | University Of Shanghai For Science And Technology | Method for identifying authenticity and origin of Panax quinquefolius based on terahertz spectroscopy |
-
2009
- 2009-06-23 CN CN 200910069378 patent/CN101685089B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| GUOXIANG XIE等: "Ultra-performance LC/TOF MS analysis of medicinal Panax herbs for metabolomic research", 《J. SEP. SCI.》 * |
| 李丽等: "高效液相色谱-电喷雾质谱联用法测定人参和西洋参的皂苷类成分", 《分析化学》 * |
| 汪江山等: "超高效液相色谱/飞行时间质谱用于人参皂甙Rg3作用后大鼠尿液代谢物指纹图谱分析及标记物的鉴定", 《色谱》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103592376A (en) * | 2013-07-02 | 2014-02-19 | 牡丹江友搏药业股份有限公司 | Novel quality control method of Shuxuetong injection |
| CN103592376B (en) * | 2013-07-02 | 2014-12-10 | 牡丹江友搏药业股份有限公司 | Novel quality control method of Shuxuetong injection |
| CN105301162A (en) * | 2015-11-10 | 2016-02-03 | 天津市尖峰天然产物研究开发有限公司 | Method for measuring pseudo-ginsenoside F11 in American ginseng through pre-column derivatization high performance liquid chromatography |
| CN106645482A (en) * | 2016-12-26 | 2017-05-10 | 河北省药品检验研究院 | Method for determining content of four ginsenoside components in 27-flavor Dingkun pill |
| CN107064329A (en) * | 2016-12-26 | 2017-08-18 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of Xueshuan xinmaining Tablet |
| CN107064328A (en) * | 2016-12-26 | 2017-08-18 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of ginseng spleen-strengthening bolus |
| CN107064328B (en) * | 2016-12-26 | 2018-09-21 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of ginseng spleen-strengthening bolus |
| CN107064329B (en) * | 2016-12-26 | 2018-09-25 | 河北省药品检验研究院 | The content assaying method of five kinds of Ginsenosides in a kind of Xueshuan xinmaining Tablet |
| CN110568096A (en) * | 2019-09-09 | 2019-12-13 | 哈尔滨工业大学（威海） | rapid detection method for multiple saponins of American ginseng |
| US11402325B2 (en) | 2021-01-15 | 2022-08-02 | University Of Shanghai For Science And Technology | Method for identifying authenticity and origin of Panax quinquefolius based on terahertz spectroscopy |
| CN114113444A (en) * | 2021-11-24 | 2022-03-01 | 天津中医药大学 | Method for identifying traditional Chinese medicine derived from Panax plant or different parts thereof and application thereof |
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