CN107132291A - A kind of ultra high efficiency liquid chromatography mass spectrometric combination determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule - Google Patents

A kind of ultra high efficiency liquid chromatography mass spectrometric combination determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule Download PDF

Info

Publication number
CN107132291A
CN107132291A CN201710387576.1A CN201710387576A CN107132291A CN 107132291 A CN107132291 A CN 107132291A CN 201710387576 A CN201710387576 A CN 201710387576A CN 107132291 A CN107132291 A CN 107132291A
Authority
CN
China
Prior art keywords
kinds
compositions
granule
evodia rutaecarpa
liquid chromatography
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201710387576.1A
Other languages
Chinese (zh)
Other versions
CN107132291B (en
Inventor
牛丽颖
王晓亚
王鑫国
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenwei Pharmaceutical Group Co Ltd
Hebei University of Chinese Medicine
Original Assignee
Hebei Medical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hebei Medical University filed Critical Hebei Medical University
Priority to CN201710387576.1A priority Critical patent/CN107132291B/en
Publication of CN107132291A publication Critical patent/CN107132291A/en
Application granted granted Critical
Publication of CN107132291B publication Critical patent/CN107132291B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

The present invention relates to the quantitative approach that a kind of combination of ultra high efficiency liquid chromatography mass spectrometric determines 10 kinds of compositions in evodia rutaecarpa granule.Its feature：Using LC-MS, by A phases of acetonitrile, 0.12% acetic acid/1mmol/L ammonium acetates be B phases, through 4 transformation of scale gradient elutions, flow velocity 0.8mL/min；40 DEG C of column temperature；Monitor negative ions under the scan mode of (MRM) with multiple reaction to monitor simultaneously, ion gun is ESI sources, atomization gas (GS1, N2), auxiliary gas (GS2, N2), whole process uses nitrogen, and collision atmospheric pressure is medium, Q1 and Q3 resolution ratio is UNIT；Determine evodia rutaecarpa granule Content of Chlorogenic Acid, caffeic acid, rutin, Hyperoside, aurantiamarin, dehydroevodiamine, limonin, rutaecarpin, Rutaecarpine and evocarpine content, the detection in 22 minutes of 10 kinds of compositions is finished, method is quick, accurate, noiseless.

Description

A kind of combination of ultra high efficiency liquid chromatography mass spectrometric determines in evodia rutaecarpa granule 10 kinds of compositions Quantitative approach
Technical field
A kind of ultra high efficiency liquid chromatography mass spectrometric combination determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule, belongs to quality Control field, i.e. Ultra Performance Liquid Chromatography instrument is together in series with mass spectrograph, liquid chromatogram carries out mixture separation, mass spectrograph Multiple components are detected and are quantitative determined, the characteristics of with high sensitivity, high selectivity, high accuracy, 10 kinds of compositions, 22 points Determine and finish in clock.No longer different by heterogeneity best detection wavelength difference, content height, polarity spectrum is greatly different to be stranded Disturb, test limit can by efficient liquid phase g grades of μ, be reduced to ng grades, significantly expand detection range.
Background technology
Evodia rutaecarpa is rutaceae evodia rutaecarpa Euodia rutaecarpa (Juss.) Benth., Shi Hu Euodia Rutaecarpa (Juss.) Benth.var.officinalis (Dode) Huang or thin hair evodia rutaecarpa Euodia Rutaecarpa (Juss.) Benth.var.bodinieri (Dode) Huang drying almost ripe fruit.It is during one kind is commonly used Medicine, tool cold dispelling is antipruritic, stopping nausea and vomiting by lowering the adverse flow of QI, effect of supporing yang antidiarrheal.For jueyin headache, cold hernia stomachache, the disturbance of lower legs due to pathogenic cold and dampness, through row abdomen Bitterly, abdominal distention, vomits acid regurgitation, Diarrhoea.It is main to contain composition of alkaloids, such as rutaecarpin, Rutaecarpine, dehydrogenation Wu Zhu The new ketone alkali of cornel alkali, evocarpine, hydroxyevodiamine, evocarpine, evodia rutaecarpa, evoxanthine etc.；More also Flavones ingredient, such as aurantiamarin, Hyperoside, Quercetin -3-O- β-D galactosides, Isorhamnetin -3-O- β-D galactosides, Anthocyanin, iso-amylene flavones etc.；Limonin, such as evodine, limonin, evodine acetic acid esters；It is organic Acids, such as chlorogenic acid, amino acid.Its quality standard has recorded Wu under Chinese Pharmacopoeia one evodia rutaecarpa medicinal material of version in 2015 The thin layer of fruit of medicinal cornel alkali and Rutaecarpine differentiates and rutaecarpin, Rutaecarpine, the efficient liquid phase quantitative determination of limonin. Mostly in current preparation is, using rutaecarpin, Rutaecarpine as index, to be quantitative determined.Although Zeng Jinxiang, Zhu Jixiao, Huang Green great waves etc. are topic with " assay of 7 kinds of compositions in evodia rutaecarpa ", it was recently reported that use high performance liquid chromatography, are eluted through 5 subgradients, Determine synephrine, chlorogenic acid, rutaecarpin, Rutaecarpine, Quercetin, rutin and Hyperoside seven kinds of compositions in evodia rutaecarpa Document, but a set of Gradient program will spend nearly 50 minutes plus initial gradient percentage is equilibrated to, and need testing solution separation Crest can not return baseline, and Interference Peaks are too many, it is difficult to accurate quantitative analysis (see Fig. 1-A, B).Yin Yuanyuan, Yan Lihua, Zhang Qiwei etc. are reported It is B phases, 5mmolL by A phases, tetrahydrofuran of acetonitrile-1Ammonium acetate buffer (pH3.8) is C phases, is eluted through 8 subgradients, Evodia rutaecarpa is determined with HPLC methods and its commonly uses N- formyls dihydro Rutaecarpine, rutaecarpin, Rutaecarpine, 1- in product medicinal material Methyl -2- undecyl -4- quinolones, evocarpine, the content method of dihydro evocarpine and limonin, but Run a pin and spend minute about 90 minutes, and separation chromatography figure baseline is uneven, it is difficult to applied in produce reality.
Do not find ultra performance liquid chromatography mass spectrometry also at present, efficiently, quick determine in evodia rutaecarpa 10 kinds of compositions Report.It is the quality of efficient, quick, accurate multi-objective control evodia rutaecarpa granule under above-mentioned background, invention The combination of ultra high efficiency liquid chromatography mass spectrometric determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule.
The preparation method of evodia rutaecarpa granule takes evodia rutaecarpa medicine materical crude slice 6000g, and extracting in water is secondary, 1 hour every time, carries 95~100 DEG C of temperature is taken, is filtered, filtrate decompression concentration, concentrate adds dextrin in right amount, spray drying obtains dry extract, adds Appropriate dextrin, is mixed, and 1000g is made in granulation, dispenses, produces.
The content of the invention
It is combined using ultra high efficiency liquid chromatography mass spectrometric, liquid chromatogram：Chromatographic column：Diamonsil C18Post (150 × 4.6mm, 5 μ m)；By A phases of acetonitrile, 0.12% acetic acid/1mmol/L ammonium acetates be B phases, using gradient elution mode, i.e. 0~6min, 15% ~28%A；6~10min, 28%~68%A；10~11min, 68%~90%A；11~22min, 90%A；Flow velocity 0.8mL/ min；40 DEG C of column temperature；Sample size is 10 μ L；Mass Spectrometry Conditions：Negative ions under the scan mode of (MRM) are monitored using multiple reaction Monitor simultaneously, ion gun is ESI sources, ion spray voltage is set to 5500 and -4500V, and source temperature (TEM) is 650 DEG C, atomization gas (GS1, N2) is 60psi, and auxiliary gas (GS2, N2) is 65psi, and interface continuous heating, whole process uses nitrogen, and gas curtain gas (N2) is set 25psi is set to, collision atmospheric pressure is medium, Q1 and Q3 resolution ratio is UNIT；The monitoring ion pair of each testing compound Molecular ion peak and daughter ion peak for compound；The holdup time (dwell time) of each ion pair is 10ms, 10 kinds into The mass spectrometry parameters divided are shown in Table 1；Data acquisition software be Analyst TF1.6 software (AB SCIEX, Foster City, CA)；
Determine evodia rutaecarpa granule Content of Chlorogenic Acid, caffeic acid, rutin, Hyperoside, aurantiamarin, dehydrogenation Wu Zhu simultaneously Cornel alkali, limonin, rutaecarpin, Rutaecarpine and evocarpine content, 10 kinds of compositions detection in 22 minutes are finished, side Method is easy, quick, accurate, noiseless.To report first.See the ion stream chromatogram (A, B in Fig. 2-11) of 10 kinds of constituents extractions And the ion scan figure (C in Fig. 2-11) of reference substance.
Pass through methodological study, the range of linearity (table 2) of 10 kinds of compositions：Chlorogenic acid sample introduction concentration is in 32.93- 3292.80ng/ml, is in good linear relationship with peak area, and regression equation is：Y=3.21e3X+4.00e4, r=0.9994； Caffeic acid sample introduction concentration, in 49.68-4968.00ng/ml, is in good linear relationship with peak area, and regression equation is：Y= 4.18e2X+2.63e4, r=0.9980；Rutin sample introduction concentration is in 5.74-573.50ng/ml, with peak area in good linear pass It is that regression equation is：Y=7.27e2X+9.22e3, r=0.9964；Hyperoside sample introduction concentration in 6.22-622.38ng/ml, It is in good linear relationship with peak area, regression equation is：Y=2.84e3X+1.41e4, r=0.9987；Aurantiamarin sample introduction concentration It is in good linear relationship with peak area in 16.63-1662.86ng/ml, regression equation is：Y=1.31e3X+4.53e4, r= 0.9994；Dehydroevodiamine sample introduction concentration is in good linear relationship with peak area, returned in 61.45-6144.60ng/ml Equation is：Y=5.58e2X+5.03e4, r=0.9988；Limonin sample introduction concentration is in 298.41-29841.00ng/ml, with peak Area is in good linear relationship, and regression equation is：Y=2.07e2X+2.17e5, r=0.9974；The sample introduction concentration of rutaecarpin It is in good linear relationship with peak area in 20.38-2037.70ng/ml, regression equation is：Y=6.88e3X+6.01e5R= 0.9954；Rutaecarpine sample introduction concentration, in 20.04-2003.90ng/ml, is in good linear relationship, recurrence side with peak area Cheng Wei：Y=2.64e4X+3.17e5, r=0.9991；The sample introduction concentration of evocarpine in 21.36-2136.40ng/ml, with Peak area is in good linear relationship, and regression equation is：Y=5.09e5X+3.78e6, r=0.9982.Reclaim real using sample-adding Test, as a result show：The average recovery rate of chlorogenic acid is 100.38% (n=6), and RSD is 1.99% (being shown in Table 3)；It is caffeinic flat The equal rate of recovery is 100.49% (n=6), and RSD is 1.38% (being shown in Table 4)；The average recovery rate of rutin is 99.67% (n=6), RSD is 2.96% (being shown in Table 5)；The average recovery rate of Hyperoside is 99.67% (n=6), and RSD is 2.07% (being shown in Table 6)；Orange The average recovery rate of skin glycosides is 100.95% (n=6), and RSD is 3.43% (being shown in Table 7)；The average recovery rate of dehydroevodiamine For 95.91% (n=6), RSD is 1.35% (being shown in Table 8)；The average recovery rate of limonin is 99.56% (n=6), and RSD is 1.70% (being shown in Table 9)；The average recovery rate of rutaecarpin is 97.12% (n=6), and RSD is 2.44% (being shown in Table 10)；Evodia rutaecarpa The average recovery rate of secondary alkali is 99.26% (n=6), and RSD is 2.93% (being shown in Table 11)；The average recovery rate of evocarpine For 99.54% (n=6), RSD is 2.28% (being shown in Table 12)；Precision (being shown in Table 13,14), stability (being shown in Table 15), repeatability The serial of methods parameter such as (being shown in Table 16), meets quantitation methodology regulation, it is adaptable to 10 kinds of compositions in evodia rutaecarpa granule Quantitative determination.
The technical solution adopted for the present invention to solve the technical problems is：
(1) testing conditions ultra performance liquid chromatography：Chromatographic column：Diamonsil C18Post (150 × 4.6mm, 5 μm)；With second Nitrile is that A phases, 0.12% acetic acid/1mmol/L ammonium acetates are B phases, using gradient elution mode, i.e. 0~6min, 15%~28%A； 6~10min, 28%~68%A；10~11min, 68%~90%A；11~22min, 90%A；Flow velocity 0.8mL/min；Column temperature 40℃；Sample size is 10 μ L；Mass Spectrometry Conditions：Negative ions under the scan mode of (MRM) are monitored using multiple reaction to monitor simultaneously, Ion gun is ESI sources, and ion spray voltage is set to 5500 and -4500V, and source temperature (TEM) is 650 DEG C, atomization gas (GS1, N2) For 60psi, auxiliary gas (GS2, N2) is 65psi, and interface continuous heating, whole process uses nitrogen, and gas curtain gas (N2) is set to 25psi, collision atmospheric pressure is medium, Q1 and Q3 resolution ratio is UNIT；The monitoring ion pair of each testing compound is change The molecular ion peak and daughter ion peak of compound；The holdup time (dwell time) of each ion pair is 10ms；Data acquisition is soft Part is Analyst TF1.6 software (AB SCIEX, Foster City, CA)；
(2) precision weighs chlorogenic acid, caffeic acid, rutin, Hyperoside, aurantiamarin, gone respectively for the preparation of reference substance solution Hydrogen rutaecarpin, limonin, rutaecarpin, Rutaecarpine and appropriate evocarpine, are dissolved and are configured to methanol Certain density storing solution.Precision measures certain volume storing solution respectively, mixing, and be diluted to concentration difference 329.28, 496.80th, 57.35,62.24,166.29,614.46,2984.10,203.77,200.39,213.64ng/mL mixing control Product solution；
(3) the preparation precision of need testing solution weighs evodia rutaecarpa granule about 0.1g, puts in conical flask, and precision is added 70% methanol 25mL, weighed weight, ultrasonically treated (power 250W, frequency 40kHz) 30min is let cool, then weighed weight, is used 70% methanol supplies the weight of less loss, shakes up, filtration, and precision measures subsequent filtrate 1ml, put in 10ml measuring bottles, plus 70% methanol is dilute Release to scale, shake up, filtered with 0.22 μm of miillpore filter, take subsequent filtrate as need testing solution；
(4) determination method is accurate respectively draws above-mentioned reference substance solution and each 10 μ l of need testing solution, injects liquid chromatogram matter Combined instrument is composed, the content of 10 kinds of compositions in need testing solution is determined.
The principle of the present invention is as follows：
10 kinds of compositions in evodia rutaecarpa granule, chemical constitution is different, and physicochemical property is different, maximum absorption wavelength and It is different, but have fixed molecular weight, as mobile phase is entered after liquid phase instrument, initial gross separation is carried out, the material of separation enters After mass spectrometer, it is changed into molecular ion and daughter fragment ion, determines ion-pair compound molecular ion peak and daughter ion peak, enter And the analysis condition of composition to be measured is determined, the peak at composition retention time to be measured is found, to integrating peak areas, its peak area is one Determine in scope and its content has good linear relationship, and be used to quantitative determine.
The present invention innovative point and have the beneficial effect that：
(1) use ultra performance liquid chromatography-mass spectrometry, by A phases of acetonitrile, 0.12% acetic acid/1mmol/L ammonium acetates be B phases, through 4 transformation of scale gradient elutions, flow velocity 0.8mL/min；40 DEG C of column temperature；The scanning side of (MRM) is monitored with multiple reaction Negative ions are monitored simultaneously under formula, and ion gun is ESI sources, and atomization gas (GS1, N2) aids in gas (GS2, N2), whole process uses nitrogen Gas, collision atmospheric pressure is medium, Q1 and Q3 resolution ratio is UNIT；Determine evodia rutaecarpa granule Content of Chlorogenic Acid, coffee Acid, rutin, Hyperoside, aurantiamarin, dehydroevodiamine, limonin, rutaecarpin, Rutaecarpine and evodia rutaecarpa card product Alkali content, 10 kinds of compositions detection in 22 minutes is finished, and method is easy, quick, accurate, noiseless.To report first.
(2) ultra performance liquid chromatography-mass spectrometry is compared with general high performance liquid chromatography, with high sensitivity, Gao Xuan The characteristics of selecting property, high accuracy, 10 kinds of compositions are determined in 22 minutes and finished.No longer by heterogeneity best detection wavelength it is different, Content height it is different, polarity spectrum great disparity puzzlement, test limit can by efficient liquid phase g grades of μ, be reduced to ng grades, significantly Expand detection range.The assay collection of illustrative plates of 7 kinds of compositions is learnt in above-mentioned evodia rutaecarpa, and the short time is difficult on liquid chromatograph With the micro constitutent for separating or quantitative determining, it is combined by liquid phase mass spectrograph, obtains good separation, obtaining can accurate quantitative analysis survey Fixed crest information.
(3) rutin, Hyperoside, aurantiamarin belong to flavones ingredient, and caffeic acid and chlorogenic acid are organic acid, and lemon is bitter Element is limonin, and rutaecarpin, Rutaecarpine, dehydroevodiamine and evocarpine are alkaloids, this A little Components Chemical structures are as follows：
Learnt by structure chart, structure is different, polarity spectrum great disparity, by the big minispread of polarity, it goes out on liquid phase mass spectrograph The order at peak is：Chlorogenic acid → caffeic acid → rutin → Hyperoside → aurantiamarin → dehydroevodiamine → limonin → Wu Fruit of medicinal cornel alkali → Rutaecarpine → evocarpine, using general efficient liquid phase, it is difficult to which it is good to obtain separation in a short time Good crest；Best detection wavelength and it is different, it is difficult to unified；Content height is different, has some in liquid phase without crest information；Institute To use common efficient liquid phase, it is impossible within 22 minutes, different structure, polarity great disparity, content height difference more than 700 are determined 10 kinds of compositions again.And ultra performance liquid chromatography mass spectrometry is used, succeed detection.For many fingers of evodia rutaecarpa granule Mark quick detection and provide method.
Brief description of the drawings
The assay HPLC chromatogram of 7 kinds of reference substances of Fig. 1-A documents
The assay HPLC chromatogram of 7 kinds of compositions in Fig. 1-B document evodia rutaecarpas
The ion stream chromatogram that Fig. 2-A chlorogenic acids reference substance solution is extracted
The ion stream chromatogram that Fig. 2-B need testing solutions are extracted
The ion scan figure of Fig. 2-C chlorogenic acid reference substance solutions
The ion stream chromatogram that Fig. 3-A caffeic acids reference substance solution is extracted
The ion stream chromatogram that Fig. 3-B need testing solutions are extracted
The ion scan figure of Fig. 3-C caffeic acid reference substance solutions
The ion stream chromatogram that Fig. 4-A control substance of Rutin solution is extracted
The ion stream chromatogram that Fig. 4-B need testing solutions are extracted
The ion scan figure of Fig. 4-C control substance of Rutin solution
The ion stream chromatogram that Fig. 5-A Hyperosides reference substance solution is extracted
The ion stream chromatogram that Fig. 5-B need testing solutions are extracted
The ion scan figure of Fig. 5-C Hyperoside reference substance solutions
The ion stream chromatogram that Fig. 6-A aurantiamarins reference substance solution is extracted
The ion stream chromatogram that Fig. 6-B need testing solutions are extracted
The ion scan figure of Fig. 6-C aurantiamarin reference substance solutions
The ion stream chromatogram that Fig. 7-A dehydroevodiamines reference substance solution is extracted
The ion stream chromatogram that Fig. 7-B need testing solutions are extracted
The ion scan figure of Fig. 7-C dehydroevodiamine reference substance solutions
The ion stream chromatogram that Fig. 8-A limonins reference substance solution is extracted
The ion stream chromatogram that Fig. 8-B need testing solutions are extracted
The ion scan figure of Fig. 8-C limonin reference substance solutions
The ion stream chromatogram that Fig. 9-A rutaecarpins reference substance solution is extracted
The ion stream chromatogram that Fig. 9-B need testing solutions are extracted
The ion scan figure of Fig. 9-C rutaecarpin reference substance solutions
The ion stream chromatogram that Figure 10-A Rutaecarpines reference substance solution is extracted
The ion stream chromatogram that Figure 10-B need testing solutions are extracted
The ion scan figure of Figure 10-C Rutaecarpine reference substance solutions
The ion stream chromatogram that Figure 11-A evocarpines reference substance solution is extracted
The ion stream chromatogram that Figure 11-B need testing solutions are extracted
The ion scan figure of Figure 11-C evocarpine reference substance solutions
In Fig. 1-A, B, ordinate is absorption intensity；Abscissa is retention time (minute), wherein, 1. synephrines 2. are green The Rutaecarpine of 6. rutaecarpin of ortho acid 3. rutin, 4. Hyperoside, 5. Quercetin 7.
A and B figures in Fig. 2~Figure 11, ordinate is absorption intensity；Abscissa be retention time (minute), wherein A for pair According to product, B is test sample.
C ordinates in Fig. 2~Figure 11 are absorption intensity；Abscissa is molecular weight, from left to right, and the 1st indicates in figure Numeral be that the daughter ion peak of each reference substance, the 2nd numeral are molecular ion peak.
The specific embodiment of the invention
Embodiment：
(1) testing conditions ultra performance liquid chromatography：Chromatographic column：Diamonsil C18Post (150 × 4.6mm, 5 μm)；With second Nitrile is that A phases, 0.12% acetic acid/1mmol/L ammonium acetates are B phases, using gradient elution mode, i.e. 0~6min, 15%~28%A； 6~10min, 28%~68%A；10~11min, 68%~90%A；11~22min, 90%A；Flow velocity 0.8mL/min；Column temperature 40℃；Sample size is 10 μ L；Mass Spectrometry Conditions：Negative ions under the scan mode of (MRM) are monitored using multiple reaction to monitor simultaneously, Ion gun is ESI sources, and ion spray voltage is set to 5500 and -4500V, and source temperature (TEM) is 650 DEG C, atomization gas (GS1, N2) For 60psi, auxiliary gas (GS2, N2) is 65psi, and interface continuous heating, whole process uses nitrogen, and gas curtain gas (N2) is set to 25psi, collision atmospheric pressure is medium, Q1 and Q3 resolution ratio is UNIT；The monitoring ion pair of each testing compound is change The molecular ion peak and daughter ion peak of compound；The holdup time (dwell time) of each ion pair is 10ms；Data acquisition is soft Part is Analyst TF1.6 software (AB SCIEX, Foster City, CA)；
(2) precision weighs chlorogenic acid, caffeic acid, rutin, Hyperoside, aurantiamarin, gone respectively for the preparation of reference substance solution Hydrogen rutaecarpin, limonin, rutaecarpin, Rutaecarpine and appropriate evocarpine, are dissolved and are configured to methanol Certain density storing solution.Precision measures certain volume storing solution respectively, mixing, and be diluted to concentration difference 329.28, 496.80th, 57.35,62.24,166.29,614.46,2984.10,203.77,200.39,213.64ng/mL mixing control Product solution；
(3) the preparation precision of need testing solution weighs evodia rutaecarpa granule about 0.1g, puts in conical flask, and precision is added 70% methanol 25mL, weighed weight, ultrasonically treated (power 250W, frequency 40kHz) 30min is let cool, then weighed weight, is used 70% methanol supplies the weight of less loss, shakes up, filtration, and precision measures subsequent filtrate 1ml, put in 10ml measuring bottles, plus 70% methanol is dilute Release to scale, shake up, filtered with 0.22 μm of miillpore filter, take subsequent filtrate as need testing solution.
(4) determination method is accurate respectively draws above-mentioned reference substance solution and each 10 μ l of need testing solution, injects liquid chromatogram matter Combined instrument is composed, the content of 10 kinds of compositions in need testing solution is determined.7 batches of samples of different manufacturers are determined, 17 are the results are shown in Table.
The mass spectrometry parameters of 1 10 kinds of tested compositions of table
The regression analysis equation and test limit, quantitative limit of the test compound of table 2
The chlorogenic acid of table 3 is loaded recovery test result
The caffeic acid of table 4 is loaded recovery test result
The rutin of table 5 is loaded recovery test result
The Hyperoside of table 6 is loaded recovery test result
The aurantiamarin of table 7 is loaded recovery test result
The dehydroevodiamine of table 8 is loaded recovery test result
The limonin of table 9 is loaded recovery test result
The rutaecarpin of table 10 is loaded recovery test result
The Rutaecarpine of table 11 is loaded recovery test result
The evocarpine of table 12 is loaded recovery test result
The withinday precision of table 13
The day to day precision of table 14
The stability of table 15
The replica test of table 16 (mg/g)
Note：Each component order of table 13~16 is the order of instrument automatic printing, is not adjusted again by peak sequence.
The content (mg/g) of 10 kinds of chemical compositions in the evodia rutaecarpa granule of table 17

Claims (3)

1. a kind of ultra high efficiency liquid chromatography mass spectrometric combination determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule, its feature exists In：
(1) testing conditions
Liquid chromatogram：Chromatographic column：Diamonsil C18Post (150 × 4.6mm, 5 μm)；By A phases of acetonitrile, 0.12% acetic acid/ 1mmol/L ammonium acetates are B phases, using gradient elution mode, i.e. 0~6min, 15%~28%A；6~10min, 28%~68% A；10~11min, 68%~90%A；11~22min, 90%A；Flow velocity 0.8mL/min；40 DEG C of column temperature；Sample size is 10 μ L；
Mass Spectrometry Conditions：Monitor negative ions under the scan mode of (MRM) using multiple reaction to monitor simultaneously, ion gun is ESI sources, Ion spray voltage is set to 5500 and -4500V, and source temperature (TEM) is 650 DEG C, and atomization gas (GS1, N2) is 60psi, aids in gas (GS2, N2) is 65psi, and interface continuous heating, whole process uses nitrogen, and gas curtain gas (N2) is set to 25psi, and collision atmospheric pressure is Medium, Q1 and Q3 resolution ratio are UNIT；The monitoring ion pair of each testing compound for compound molecular ion peak with Daughter ion peak；The holdup time (dwell time) of each ion pair is 10ms；Data acquisition software is Analyst TF 1.6 Software (AB SCIEX, Foster City, CA)；
(2) precision weighs chlorogenic acid, caffeic acid, rutin, Hyperoside, aurantiamarin, dehydrogenation Wu respectively for the preparation of reference substance solution Fruit of medicinal cornel alkali, limonin, rutaecarpin, Rutaecarpine and appropriate evocarpine, are dissolved with methanol and are configured to certain The storing solution of concentration.Precision measures certain volume storing solution respectively, mixing, and be diluted to concentration difference 329.28,496.80, 57.35th, 62.24,166.29,614.46,2984.10,203.77,200.39,213.64ng/mL mixed reference substance solution；
(3) the preparation precision of need testing solution weighs evodia rutaecarpa granule about 0.1g, puts in conical flask, and precision adds 70% Methanol 25mL, weighed weight, ultrasonically treated (power 250W, frequency 40kHz) 30min is let cool, then weighed weight, uses 70% first Alcohol supplies the weight of less loss, shakes up, and filtration, precision measures subsequent filtrate 1ml, put in 10ml measuring bottles, plus 70% methanol dilution is extremely carved Degree, shakes up, and is filtered with 0.22 μm of miillpore filter, takes subsequent filtrate as need testing solution.
(4) determination method is accurate respectively draws above-mentioned reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography mass connection With instrument, the content of 10 kinds of compositions in need testing solution is determined.
2. a kind of ultra high efficiency liquid chromatography mass spectrometric combination according to claim 1 determines in evodia rutaecarpa granule 10 kinds of compositions Quantitative approach, it is characterised in that 10 kinds of described compositions are chlorogenic acid, caffeic acid, rutin, Hyperoside, aurantiamarin, dehydrogenation Wu Fruit of medicinal cornel alkali, limonin, rutaecarpin, Rutaecarpine and evocarpine.
3. a kind of ultra high efficiency liquid chromatography mass spectrometric combination according to claim 1 determines in evodia rutaecarpa granule 10 kinds of compositions Quantitative approach, it is characterised in that described evodia rutaecarpa granule is per the suitable primary crude drug 6g of 1g.
CN201710387576.1A 2017-05-22 2017-05-22 Quantitative method for measuring 10 components in evodia rutaecarpa formula particles by ultra-performance liquid mass spectrometry Active CN107132291B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710387576.1A CN107132291B (en) 2017-05-22 2017-05-22 Quantitative method for measuring 10 components in evodia rutaecarpa formula particles by ultra-performance liquid mass spectrometry

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710387576.1A CN107132291B (en) 2017-05-22 2017-05-22 Quantitative method for measuring 10 components in evodia rutaecarpa formula particles by ultra-performance liquid mass spectrometry

Publications (2)

Publication Number Publication Date
CN107132291A true CN107132291A (en) 2017-09-05
CN107132291B CN107132291B (en) 2020-05-19

Family

ID=59735045

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710387576.1A Active CN107132291B (en) 2017-05-22 2017-05-22 Quantitative method for measuring 10 components in evodia rutaecarpa formula particles by ultra-performance liquid mass spectrometry

Country Status (1)

Country Link
CN (1) CN107132291B (en)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107843674A (en) * 2017-12-26 2018-03-27 天圣制药集团股份有限公司 The detection method of active component in a kind of evodia rutaecarpa juice
CN108037197A (en) * 2017-11-27 2018-05-15 中山市中智药业集团有限公司 A kind of method of the non-volatile class secondary metabolite chemical composition Qualitative and quantitative analysis of cordate houttuynia broken wall medicine materical crude slice
CN108195957A (en) * 2017-12-19 2018-06-22 江苏省农业科学院 The high performance liquid chromatography tandem mass spectrum detection method of phenolic acid in a kind of wheat being simple and efficient
CN109212120A (en) * 2018-09-30 2019-01-15 湖南省中医药研究院 A kind of construction method and evodia rutaecarpa quality of medicinal material detection method of evodia rutaecarpa medicinal material characteristic spectrum
CN109568373A (en) * 2019-01-24 2019-04-05 广东方制药有限公司 A method of pulp of dogwood fruit or wine-prepared fructus corni granule are prepared using continuous dynamic countercurrent extraction process
CN110187027A (en) * 2019-05-30 2019-08-30 贵州医科大学 A kind of method of quality control of capsule preparations that treating gastric ulcer
CN110568094A (en) * 2019-08-29 2019-12-13 梅州广药采芝林药业有限公司 detection method of evodiamine, rutaecarpine and limonin in soaked fructus evodiae
CN112014480A (en) * 2019-05-28 2020-12-01 黄河科技学院 Method for detecting content of effective components in Jiangzhining granules by UPLC-MS/MS (ultra performance liquid chromatography-Mass Spectrometry/Mass Spectrometry)
CN113406225A (en) * 2021-06-11 2021-09-17 上海上药杏灵科技药业股份有限公司 Multi-component analysis method for standing circle
CN114487176A (en) * 2022-01-18 2022-05-13 大连海洋大学 Rapid screening method for dangerous substances in aquatic products
CN114755328A (en) * 2022-04-01 2022-07-15 四川新绿色药业科技发展有限公司 Method for constructing Evodia rutaecarpa high performance liquid phase characteristic spectrum and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102141555A (en) * 2010-12-27 2011-08-03 贵阳中医学院 Method for establishing high-pressure liquid-phase fingerprinting of evodia and extractives thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102141555A (en) * 2010-12-27 2011-08-03 贵阳中医学院 Method for establishing high-pressure liquid-phase fingerprinting of evodia and extractives thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
MENG-YAO ZHAO 等: "Optimization of the extraction conditions and simultaneous quantification by RP－LC of six alkaloids in Evodiae Fructus", 《CHROMATOGRAPHIA》 *
孙健 等: "高效液相色谱串联质谱分析吴茱萸水煎液的化学成分", 《中国药物警戒》 *
张建锋 等: "黔产吴茱萸的UPLC-MS/MS指纹图谱分析", 《贵州农业科学》 *
高鹏 等: "吴茱萸碱和吴茱萸次碱电喷雾-离子阱质谱裂解途径研究", 《药物分析杂志》 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108037197A (en) * 2017-11-27 2018-05-15 中山市中智药业集团有限公司 A kind of method of the non-volatile class secondary metabolite chemical composition Qualitative and quantitative analysis of cordate houttuynia broken wall medicine materical crude slice
CN108195957A (en) * 2017-12-19 2018-06-22 江苏省农业科学院 The high performance liquid chromatography tandem mass spectrum detection method of phenolic acid in a kind of wheat being simple and efficient
CN107843674A (en) * 2017-12-26 2018-03-27 天圣制药集团股份有限公司 The detection method of active component in a kind of evodia rutaecarpa juice
CN109212120B (en) * 2018-09-30 2021-10-26 湖南省中医药研究院 Method for constructing characteristic spectrum of medicinal evodia fruit and method for detecting quality of medicinal evodia fruit
CN109212120A (en) * 2018-09-30 2019-01-15 湖南省中医药研究院 A kind of construction method and evodia rutaecarpa quality of medicinal material detection method of evodia rutaecarpa medicinal material characteristic spectrum
CN109568373A (en) * 2019-01-24 2019-04-05 广东方制药有限公司 A method of pulp of dogwood fruit or wine-prepared fructus corni granule are prepared using continuous dynamic countercurrent extraction process
CN112014480A (en) * 2019-05-28 2020-12-01 黄河科技学院 Method for detecting content of effective components in Jiangzhining granules by UPLC-MS/MS (ultra performance liquid chromatography-Mass Spectrometry/Mass Spectrometry)
CN110187027A (en) * 2019-05-30 2019-08-30 贵州医科大学 A kind of method of quality control of capsule preparations that treating gastric ulcer
CN110568094A (en) * 2019-08-29 2019-12-13 梅州广药采芝林药业有限公司 detection method of evodiamine, rutaecarpine and limonin in soaked fructus evodiae
CN113406225A (en) * 2021-06-11 2021-09-17 上海上药杏灵科技药业股份有限公司 Multi-component analysis method for standing circle
CN113406225B (en) * 2021-06-11 2023-03-10 上海上药杏灵科技药业股份有限公司 Multi-component analysis method for standing circle
CN114487176A (en) * 2022-01-18 2022-05-13 大连海洋大学 Rapid screening method for dangerous substances in aquatic products
CN114487176B (en) * 2022-01-18 2023-06-16 大连海洋大学 Rapid screening method for dangerous substances in aquatic products
CN114755328A (en) * 2022-04-01 2022-07-15 四川新绿色药业科技发展有限公司 Method for constructing Evodia rutaecarpa high performance liquid phase characteristic spectrum and application thereof
CN114755328B (en) * 2022-04-01 2023-06-23 四川新绿色药业科技发展有限公司 Method for constructing high-efficiency liquid-phase characteristic spectrum of salted evodia rutaecarpa and application of method

Also Published As

Publication number Publication date
CN107132291B (en) 2020-05-19

Similar Documents

Publication Publication Date Title
CN107132291A (en) A kind of ultra high efficiency liquid chromatography mass spectrometric combination determines the quantitative approach of 10 kinds of compositions in evodia rutaecarpa granule
CN110554108B (en) Quality detection method for lindley eupatorium herb
CN109633001B (en) Fingerprint detection method of wine glossy privet fruit
CN108181400B (en) Method for simultaneously measuring contents of 6 chemical components in spina date seed water decoction by UHP L C-QQQ-MS/MS
Zhao et al. Simultaneous quantification of seven bioactive flavonoids in Citri Reticulatae Pericarpium by ultra‐fast liquid chromatography coupled with tandem mass spectrometry
CN106918667B (en) Pressurized micro-extraction equipment, pressurized micro-extraction method and application thereof
CN111751465B (en) Rapid quantitative screening method and application of liquorice antioxidant active ingredients
CN107843657A (en) The quality determining method of TONGXIAO BIYAN PIAN
CN107529337A (en) The HPLC analyses of impurity in two to the water wei ling alcohol
CN109374785B (en) Construction method and detection method of UPLC (ultra performance liquid chromatography) characteristic spectrum of lophatherum gracile medicinal material
CN108152399A (en) A kind of structure and its detection method of seed of Oriental arborvitae medicinal material UPLC characteristic spectrums
Wei et al. Quantitative analysis of sesquiterpenes and comparison of three Curcuma wenyujin herbal medicines by micro matrix solid phase dispersion coupled with MEEKC
CN110441413A (en) The construction method and detection method of qianbai biyan tablets HPLC finger-print
CN108845062B (en) High performance liquid chromatography for determining component content of compound giant knotweed rhizome ammonia sensitive tablet
Feng et al. Study on chemical constituents and fingerprints of Phellodendron amurense Rupr. based on ultra‐performance liquid chromatography–quadrupole–time‐of‐flight–mass spectrometry
CN109632995A (en) A kind of method for building up of semen ziziphi spinosae flavones ingredient UPLC finger-print and application
Zhao et al. Quantitative analysis of five toxic alkaloids in Aconitum pendulum using ultra-performance convergence chromatography (UPC 2) coupled with mass spectrometry
CN110146611B (en) Method for rapidly identifying chemical components in donkey-hide gelatin blood-enriching particles
Zhao et al. pH‐Zone‐refining counter‐current chromatography for two new lipo‐alkaloids separated from refined alkaline extraction of Kusnezoff monkshood root
CN110940755A (en) Method for detecting 4 tadalafil substances in food
CN109374788A (en) The UPLC characteristic spectrum construction method and detection method of Fructus cnidii
CN111175416B (en) Method for simultaneously detecting 7 components in dogwood
CN110346463A (en) A kind of method for building up of Flos Ilicis Asprellae root herb HPLC-ELSD finger-print
CN107383032A (en) For the morphine base of forensic science illicit drugs inspection, morphine hydrochloride, heroin hydrochloride standard substance method for preparing purified
Wang et al. Separation and purification of five alkaloids from Aconitum duclouxii by counter‐current chromatography

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20201029

Address after: 050200 No. 3 Xingyuan Road, Luquan Economic Development Zone, Shijiazhuang City, Hebei Province

Patentee after: HEBEI University OF CHINESE MEDICINE

Patentee after: Shineway Pharmaceutical Group Ltd.

Address before: 050200 No. 3 Apricot Garden Road, Luquan Economic Development Zone, Hebei, Shijiazhuang

Patentee before: HEBEI University OF CHINESE MEDICINE