CN106872616A - The method for distinguishing Dali Paris polyphylla and Yunnan Rhizoma Paridis - Google Patents
The method for distinguishing Dali Paris polyphylla and Yunnan Rhizoma Paridis Download PDFInfo
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- CN106872616A CN106872616A CN201710188767.5A CN201710188767A CN106872616A CN 106872616 A CN106872616 A CN 106872616A CN 201710188767 A CN201710188767 A CN 201710188767A CN 106872616 A CN106872616 A CN 106872616A
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
Method the invention provides Dali Paris polyphylla and Yunnan Rhizoma Paridis is distinguished, it is characterised in that：It is detected using liquid chromatography, if detecting containing chonglou saponin VII, H, 0.010% is below without Gracillin, chonglou saponin I, II or content, then can determine that to be Dali Paris polyphylla.Above-mentioned detection method, simple, accurate, analysis time is short, good stability, can be used for the differentiation of Dali Paris polyphylla and Yunnan Rhizoma Paridis.
Description
Technical field
The present invention relates to Chinese medicine detection field, and in particular to the method for distinguishing Dali Paris polyphylla and Yunnan Rhizoma Paridis.
Background technology
《Pharmacopoeia of People's Republic of China》Version one records Paris polyphylla kind for liliaceous plant Yunnan Rhizoma Paridis Paris within 2015
Polyphylla Smith var.yunnanensis (Franch.) Hand.-Mazz. or paris polyphylla Paris
The dry rhizome of polyphylla Smith var.chinensis (Franch.) Hara, with clearing heat and detoxicating, swelling and pain relieving,
Effect of cool liver arresting convulsion.Modern study shows that there is Paris polyphylla antitumor, hemostasis, immunological regulation, calmness, analgesia etc. to act on, steroidal
Saponin constituent is the main active for playing a role.In recent years, because the Paris polyphylla market demand is huge, wild resource is in imminent danger and artificial
Raising technology requirement is high, cultivation period is long etc. it is multifactor cause Paris polyphylla substantial appreciation of prices, 50 from before 10 years yuan/kg increases to 700
~900 yuan/kg.
Dali Paris polyphylla Paris daliensis H.Li et V.G.Souku and Dulong Paris polyphylla Paris dulongensis
H.Li et S.Kuritap are subordinate to Liliaceae Liliacceae Paris Paris, and rhizome is among the people in various regions such as Yunnan, Guizhou
Used as Paris polyphylla medicinal material, bitter, cold nature is slightly poisonous, the effect of arresting convulsion with clearing heat and detoxicating, swelling and pain relieving, cool liver, be used for
The diseases such as ulcerative carbuncle, abscess of throat, venomous snake bite, traumatic injury, cool breeze tic.Not yet it is indexed to version in 2015 at present《Chinese Pharmacopoeia》
Or in each provincial medicinal material standard.Due to Yunnan Baiyao as representative etc. medicine series to the demand day of Paris polyphylla class rhizome increasingly
Greatly, the rhizome of many plants of Paris, including Dali Paris polyphylla and Dulong Paris polyphylla are largely purchased as Paris polyphylla medicinal material, and Dali
Paris polyphylla and Dulong Paris polyphylla could as the source of Paris polyphylla medicinal material and their resources reserve how, the market demand could be met
Research report is little, therefore, the relationship kind of necessary Yunnan Rhizoma Paridis carries out basic research, for MED SUP provides foundation.
Yet there are no can accurately distinguish both methods in the melange of Dali Paris polyphylla and Yunnan Rhizoma Paridis.Certainly,
Also the method for distinguishing Dali Paris polyphylla and Dulong Paris polyphylla is had no.
The content of the invention
Version in 2015《Chinese Pharmacopoeia》In, with rhizoma paridis saponin I, chonglou saponin Ⅱ, polyphyllin Ⅵ and chonglou saponin VII
Mass fraction is also recorded as Paris polyphylla quality index is evaluated in patent application 2016106653439, is mainly contained in Yunnan Rhizoma Paridis
Chonglou saponin VII, H, II, I, Gracillin can also occur in the Yunnan Rhizoma Paridis of part, however, Dali Paris polyphylla is but containing only weight
Building saponin(e VII and H, do not there is chonglou saponin II, I and very thin pennogenin.Therefore, compared with Yunnan Rhizoma Paridis, contained by the Paris polyphylla of Dali
Mainly there are chonglou saponin VII and H in chemical composition, and do not contain chonglou saponin II, I and very thin pennogenin, therefore, it can lead to
Mentioned component is crossed to separate Dali Paris polyphylla with Yunnan Rhizoma Paridis discriminating.
Specifically, the invention provides the method for distinguishing Dali Paris polyphylla and Yunnan Rhizoma Paridis, it is examined using liquid chromatography
Survey, if detecting containing chonglou saponin VII, H, be below without Gracillin, chonglou saponin I, II or content
0.010%, then can determine that to be Dali Paris polyphylla, you can with the melange of Dali Paris polyphylla and Yunnan Rhizoma Paridis, by the above method
Dali Paris polyphylla is identified, the effect of differentiation is reached.
Further, wherein, the content of chonglou saponin VII is more than 0.3%；The content of chonglou saponin H 0.3% with
On.
Further, wherein, the content without chonglou saponin VI or chonglou saponin VI be less than 0.02%.
Further, wherein, the total content of chonglou saponin VII, VI, II and I is more than 0.3%；Chonglou saponin VII, VI,
The total content of II, I, H and Gracillin is more than 1.4%.
Wherein, the liquid chromatography is selected from UPLC, and chromatographic condition of the present invention can be detected using known method, such as
Patent application 2016106653439, it is also possible to carried out according to following conditions of the invention：The concrete operations of the UPLC are as follows：
(1) dry measuring samples are taken, is crushed, with alcohols solvent or aqueous alcohols solvent extraction, merge extract solution, removed
Remaining residue methyl alcohol after solvent, aqueous methanol, acetonitrile redissolve containing water-acetonitrile, prepare need testing solution；
(2) rhizoma paridis saponin I, II, VI, VII, H, the mixture of Gracillin are taken, reference substance solution is prepared；
(3) need testing solution, reference substance solution are injected separately into Ultra Performance Liquid Chromatography instrument, are using external standard method detection
Can；Wherein, chromatographic condition is as follows：
Chromatographic column：Octadecylsilane chemically bonded silica is filler,
ELSD detector conditions：55 ± 2 DEG C of drift tube temperature, 40 ± 2psi of nitrogen pressure；
Mobile phase：Acetonitrile (A)-water (B) system, its Gradient program is as follows：
0~1min, (5% or 10%)~(25% or 30%) A；1~(2.5 or 3) min, (25% or 30%)~30%
A；(2.5 or 3)~4min, 30%~(35% or 40%) A；4~(6 or 8) min, (35% or 40%)~40%A；(6 or 8)
~20min, 40%~50%A；20~22min, 50%~70%A；22-23min, 70%~90%A.
Wherein, in step (1), alcohols solvent be selected from methyl alcohol, ethanol or propyl alcohol, aqueous alcohols solvent be selected from aqueous methanol,
Hydrous ethanol or aqueous propyl alcohol；In step (2), the solvent of reference substance solution is selected from methyl alcohol, aqueous methanol, acetonitrile or aqueous second
Nitrile.
Wherein, in step (3), described column size model etc. should meet the requirement of UPLC, and the present invention one is specific real
Apply in mode, column size is 2.1 × 50mm, and the particle diameter of filler is 1.8 μm.
Wherein, 0.1~0.3mLmin of flow rate of mobile phase-1；40 ± 2 DEG C of column temperature.Certainly, determine in condition of gradient elution
In the case of, appropriate change flow velocity and column temperature can equally obtain satisfied measurement result, but the retention time at each peak can accordingly change
Become, can be determined by reference substance chromatographic peak.
Wherein, eluent gradient is：
0~1min, 10%~30%A；1~3min, 30%~30%A；3~4min, 30%~40%A；4~8min,
40%~40%A；8~20min, 40%~50%A；20~22min, 50%~70%A；22~23min, 70%~90%A.
Method present invention also offers Dali Paris polyphylla and Dulong Paris polyphylla is distinguished, it is detected using liquid chromatography, if
Detect containing chonglou saponin VII, H, 0.010% is below without Gracillin, chonglou saponin I, II or content, then may be used
It is judged to Dali Paris polyphylla；If detecting containing Gracillin, it is below without chonglou saponin VII, H, VI or content
0.010%, then can determine that to be Dulong Paris polyphylla.I.e. can in the melange of Dali Paris polyphylla and Dulong Paris polyphylla, by Dali Paris polyphylla and
Dulong Paris polyphylla is accurately distinguished.
Above-mentioned UPLC methods and chromatographic condition can also be used in the detection method.
Wherein, solvent used in " redissolution " can be selected from methyl alcohol, aqueous methanol, acetonitrile or containing water-acetonitrile.The present invention one
Methyl alcohol is used in individual specific embodiment as solvent.But, mobile phase should equally have good molten to target component to be checked
Solution performance, therefore, in actual use, can equally use mobile phase solvent --- acetonitrile is answered containing water-acetonitrile
It is molten.
Wherein, extracting mode is selected from ultrasound, dipping, diacolation or backflow.In a specific embodiment of the invention, from super
Sound is used as extracting mode.Certainly, which kind of extracting mode no matter the present invention select, and is all not influence target in testing sample to try one's best
Premised on the accuracy in detection of composition, in actual mechanical process, can be by identical detection method of the present invention come to extracting
Mode carries out examination, and its examination experiment is common confirmatory experiment, without involving an inventive effort.
In a specific embodiment of the invention, in step (1), solvent is removed by the way of being evaporated under reduced pressure.Wherein,
The use of " reduction vaporization ", be also based on not influenceing the accuracy in detection of target component in testing sample as far as possible for the purpose of, certainly,
Except this method, other solvent removal means, such as natural evaporation, low-temperature heat evaporation can also be used.That is, remove molten
The mode of agent is the conventional operational means in this area, can be selected according to actual conditions, be should not necessarily be limited by " reduction vaporization ".
Brief description of the drawings
Fig. 1 mixing reference substance chromatograms, 1. chonglou saponin VII；2. chonglou saponin H；3. polyphyllin Ⅵ；4. chonglou saponin
Ⅱ；5. Gracillin；6. rhizoma paridis saponin I
Fig. 2 Dali Paris polyphylla chromatogram, 1. chonglou saponin VII；2. chonglou saponin H；3. polyphyllin Ⅵ
Fig. 3 Dulong Paris polyphylla chromatograms, 1. Gracillin；2. rhizoma paridis saponin I
Fig. 4 Dali Paris polyphylla UPLC chromatograms, 1. chonglou saponin VII；2. chonglou saponin H
Fig. 5 Dulong Paris polyphylla UPLC chromatograms, 1. chonglou saponin II；2. Gracillin；3. chonglou saponin I
Specific embodiment
Unless otherwise defined herein, the scientific and technical terminology being otherwise used in conjunction with the invention should have general technology person generally to manage
The implication of solution.The implication and category of term should be clear and definite；If provided in this article however, there is any latent ambiguity
Definition is prior to any dictionary or external definition.
Embodiment 1
Instrument and material
1.1 instruments
METTLER AE240 electronic analytical balances (Shanghai plum Teller-support benefit Instrument Ltd.)；KQ-5200E types surpass
Sound wave washer (40kHz, 250W, Kunshan ultrasonic instrument Co., Ltd)；(German Sartorius is public for BP211D types electronic balance
Department)；Waters AcquityH-Class Ultra Performance Liquid Chromatography instruments (Waters, US).
1.2 materials and reagent
Water (Watson distilled water)；Methyl alcohol (analysis of Chengdu Ke Long chemical reagents factory is pure)；The HPLC grades of acetonitrile (U.S.
Sigma companies).
Compound 1：(the lot number of chonglou saponin VII：MUST-14060302, Purity：99.03%), compound 4：Paris polyphylla soap
(the lot number of glycosides VI：MUST-14071904, Purity：98.67%), compound 5：Chonglou saponin Ⅱ (lot number：MUST-
15060810, Purity：98.45%), compound 6：Gracillin (lot number：MUST-15081110, Purity：
98.86%), compound 7：Rhizoma paridis saponin I (lot number：MUST-15061611, Purity：98.34%) it is purchased from Chengdu Man Site
Bio tech ltd；(the pennogenin -3-O- β-D- glucopyranosyls (1 → 3)-[α-L- rhamnopyranosyloxyhies of compound 2
Glycosyl (1 → 2)]-β-D- glucopyranosides) and compound 3 (chonglou saponin H) be HuaXi college of pharmacy, SiChuan University Zhang Hao professor
Seminar is separating obtained, and HPLC detections purity is >=98%.
Dali Paris polyphylla and Dulong Paris polyphylla are taught by Southwest University for Nationalities national medicine research institute Liu Yuan, Sichuan University's West China medicine
Institute professor Zhang Hao identifies that former plant specimen and medicinal material Sample storage are in Southwest University for Nationalities, HuaXi college of pharmacy, SiChuan University.Sample
Product information is shown in Table 1.
Table 1
2 test methods
2.1 chromatographic conditions
ACQUITY BEH C18 chromatographic columns (2.1mm × 50mm, 1.7 μm)；55 DEG C of ELSD detectors drift tube temperature, nitrogen
Atmospheric pressure 40psi, gain 500, refrigerating mode；40 DEG C of column temperature；Flow velocity 0.2mLmin-1；Sample size：1μL；Mobile phase is acetonitrile
(A)-water (B), gradient elution program is：0-1min 10%-30%A；1-3min, 30%-30%A；3-4min, 30%-40%
A；4-8min, 40%-40%A；8-20min, 40%A-50%A；20-22min, 50%A-70%A；22-23min, 70%A-
90%A.Mixing reference substance, sample are shown in Fig. 1,2,3.
The preparation of 2.2 reference substance solutions
Respectively precision weigh chonglou saponin VII 2.060mg, chonglou saponin H 2.100mg, chonglou saponin VI 2.060mg,
Chonglou saponin II 2.080mg, Gracillin 2.040mg, chonglou saponin I 2.020mg, add methyl alcohol dissolving and constant volume respectively
It is standby through 0.22 μm of filtering with microporous membrane in 10mL volumetric flasks.Compound 2 is the great teacher of HuaXi college of pharmacy, SiChuan University
Seminar makes by oneself, and lazy weight is enough in quantitative analysis, is only used for qualitative analysis.
The preparation of 2.3 need testing solutions
Dried medicinal material powder beater beats powder, crosses 40 mesh sieves, takes medicinal material powder 0.5g, and precision weighing is placed in 100mL conical flasks
In, by solid-liquid ratio 1:100(g:ML methyl alcohol) is added, is shaken up, after standing 4h, ultrasonic 30min, filtering takes filtrate and is placed in flask, weight
Multiple aforesaid operations 2 times.Merging filtrate, is recovered under reduced pressure solution, and residue adds methyl alcohol to dissolve and be settled in 25mL volumetric flasks, is placed in 4
DEG C Refrigerator store, it is standby.0.22 μm of membrane filtration (before sample introduction) is standby.
2.4 methodological studies
2.4.1 the investigation of linear relationship
Reference substance solution is diluted to 2mL in right amount under precision draws " 2.2 " item, is made the mixing control of different extension rates
Product solution, is determined by chromatographic condition under " 2.1 " item.With peak area as ordinate (Y), sample size (μ g) is abscissa (X), is drawn
External standard method standard curve, the results are shown in Table 2.
The equation of linear regression of the reference substance of table 2
2.4.2 precision test
S3 batch need testing solutions are taken, by chromatographic condition continuous sample introduction under " 2.1 " item 6 times, record chonglou saponin VII, weight
When building saponin(e H, chonglou saponin VI, chonglou saponin II, Gracillin, chonglou saponin I, the relative reservation at chonglou saponin V peaks
Between and relative peak area.Result obtains chonglou saponin VII, VI, II, Gracillin, the RSD of I peak areas and is respectively
1.25%th, 1.33%, 1.89%, 1.67%, 1.99%, respectively less than 3%, show that instrument precision is good.
2.4.3 replica test
S3 batch need testing solutions are taken, by 6 parts of need testing solutions of parallel preparation under " 2.3 " item, by chromatostrip under " 2.1 " item
Part is measured, record chonglou saponin VII, chonglou saponin H, chonglou saponin VI, chonglou saponin II, Gracillin, Paris polyphylla
The relative retention time and relative peak area of saponin I, chonglou saponin V peaks.Result obtains chonglou saponin VII, VI, II, very thin potato
Chinese yam saponin(e, the RSD of I peak areas are respectively 1.26%, 1.21%, 1.83%, 1.27%, 1.55%, respectively less than 3%, the side of showing
Method repeatability is good.
2.4.4 stability test
S3 batch need testing solutions are taken, by chromatographic condition under " 2.1 " item, are determined respectively at 0,2,4,8,12,24h sample introductions,
Record chonglou saponin VII, chonglou saponin H, chonglou saponin VI, chonglou saponin II, Gracillin, chonglou saponin I, Paris polyphylla soap
The relative retention time and relative peak area at glycosides V peaks.Result obtains chonglou saponin VII, VI, II, Gracillin, I peaks face
Long-pending RSD is respectively 1.15%, 1.78%, 1.81%, 1.07%, 1.79%, respectively less than 3%, shows need testing solution in 24h
Internal stability is good.
2.4.5 it is loaded recovery test
Take 5 parts of the Paris polyphylla medicinal material sample of S3 batch known contents, respectively plus precision weighing add chonglou saponin VII, VI,
II, Gracillin, I reference substances, by step operation sample preparation under " 2.3 " item, constant volume is determined, and calculates the rate of recovery.Chonglou saponin
VII, VI, II, Gracillin, I mean sample recovery rates are respectively：98.92%th, 99.44%, 102.13%,
98.44%th, 99.21%, RSD is 1.86%, 1.54%, 1.48%, 1.92%, 1.83%.
3 results and analysis
Precision draws prepared need testing solution under " 2.3 " item, is determined by chromatographic condition under " 2.1 " item, calculates weight in sample
Building saponin content, Dali chonglou saponin content results are shown in Table 2, Dulong chonglou saponin content results and are shown in Table 3.Its Dali Paris polyphylla chromatogram
Figure is shown in that 4, Dulong Paris polyphylla chromatogram is shown in 5.
6 contents of steroid saponin (%) in 3 10 batches of Dali Paris polyphylla samples of table
Note："-" indicate without
6 contents of steroid saponin (%) in 47 batches of Dulong Paris polyphylla samples of table
Note："-" indicate without
Show that Paris polyphylla steroid saponin is that Paris polyphylla plays the topmost active component of pharmacological effect, steroidal in modern pharmacological research
The structure of saponin(e is similar, and end absorption is belonged on ultraviolet spectra, and sensitivity is relatively small.EISD (ELSD)
It is a kind of novel universal property amount detector for starting application the nineties in last century, it is to the compound without UV absorption in ELSD
Response signal can be produced, have the advantages that to stablize it is sensitive, can gradient elution analysis.This experiment is combined using Ultra Performance Liquid Chromatography
EISD (UPLC-ELSD) detection technique is carried out to 6 steroid saponins in Dali Paris polyphylla and Dulong Paris polyphylla medicinal material
Analysis.Result shows that can be separated 6 kinds of chonglou saponins in 24min, the method highly shortened the separation of Paris polyphylla class medicinal material
The time of analysis, the precision of method, stability, repeatability and sample-adding recovery test etc. show that institute's construction method is reliable and stable.
Dali Paris polyphylla belongs to critically endangered Paris polyphylla kind, goes through version《Chinese Pharmacopoeia》With rhizoma paridis saponin I, chonglou saponin Ⅱ,
The mass fraction total amount of polyphyllin Ⅵ and chonglou saponin VII is qualified as Paris polyphylla quality index, Zong Liang≤0.6% is evaluated.Examination
Result is tested to show：(1) the 10 batch of chonglou saponin content average value of the States Pharmacopoeia specifications of Dali Paris polyphylla is 1.2153%, except S7 batches,
Other batch Dali Paris polyphylla reach the standard of States Pharmacopoeia specifications；(2) Dali Paris polyphylla mainly contains chonglou saponin VII and chonglou saponin
H；Micro polyphyllin Ⅵ is checked in 10 batches only S10 batches, rhizoma paridis saponin I, chonglou saponin Ⅱ is not detected
With very thin pennogenin；(3) this result of the test is basically identical with the content results of seminar's early stage Dali Paris polyphylla.
Dulong Paris polyphylla is only distributed in Nujiang Prefecture Dulong River township, and its resource reserves is more rare than Dali Paris polyphylla.This is special
Journey field goes to Nujiang Prefecture Dulong River township to carry out collection of resources work, but has only collected 7 parts of samples.Result of the test shows：(1)
In 7 batches of Dulong Paris polyphylla, the mass fraction average value of rhizoma paridis saponin I, chonglou saponin Ⅱ, polyphyllin Ⅵ and chonglou saponin VII is
0.6872%, and only S13, S14, S17 batch reach the standard of the saponin content of States Pharmacopoeia specifications；(2) Dulong Paris polyphylla contains
Rhizoma paridis saponin I and very thin pennogenin, S17 batches contain chonglou saponin Ⅱ, and 7 batches are free of polyphyllin Ⅵ, Paris polyphylla soap
Glycosides VII and chonglou saponin H.
And chonglou saponin VII, H, II, I are mainly contained described in patent application 2016106653439, in Yunnan Rhizoma Paridis, and
Gracillin is not the composition certainly existed in Yunnan Rhizoma Paridis, and only 6 batches contain in 23 batches of medicinal materials, and content all exists
0.5 or lower.Therefore, compared with Yunnan Rhizoma Paridis, mainly there are rhizoma paridis saponin I and very thin Chinese yam in chemical composition contained by Dulong Paris polyphylla
Saponin(e, and polyphyllin Ⅵ, chonglou saponin VII and chonglou saponin H are not contained, meanwhile, Dali Paris polyphylla and paris polyphylla also do not have
The features described above of standby Dulong Paris polyphylla, therefore, it can differentiate separately Yunnan Rhizoma Paridis and Dulong Paris polyphylla by mentioned component.
It is attached：The content detection of paris polyphylla
Chromatographic condition
HSS C18 chromatographic columns (2.1mm × 75mm, 1.7 μm)；ELSD detector drift tube temperature
55 DEG C of degree, nitrogen pressure 40psi, gain 500, refrigerating mode；40 DEG C of column temperature；Flow velocity 0.2mLmin-1；Sample size：2μL；Stream
Dynamic is mutually acetonitrile (A)-water (B), and gradient elution program is shown in Table 5.
The gradient elution program of table 5
Each component content in the paris polyphylla of table 6
Claims (10)
1. the method for distinguishing Dali Paris polyphylla and Yunnan Rhizoma Paridis, it is characterised in that：It is detected using liquid chromatography, if detecting
Containing chonglou saponin VII, H, 0.010% is below without Gracillin, chonglou saponin I, II or content, then can determine that for
Dali Paris polyphylla.
2. differentiating method according to claim 1, it is characterised in that：Wherein, the content of chonglou saponin VII 0.3% with
On；The content of chonglou saponin H is more than 0.3%.
3. differentiating method according to claim 1, it is characterised in that：Wherein, without chonglou saponin VI or chonglou saponin
The content of VI is less than 0.02%.
4. differentiating method according to claim 1, it is characterised in that：Wherein, chonglou saponin VII, VI, II and I always contains
Amount is more than 0.3%；The total content of chonglou saponin VII, VI, II, I, H and Gracillin is more than 1.4%.
5. the differentiating method according to Claims 1 to 4 any one, it is characterised in that：The liquid chromatography is selected from
The concrete operations of UPLC, UPLC are as follows：
(1) dry measuring samples are taken, is crushed, with alcohols solvent or aqueous alcohols solvent extraction, merge extract solution, remove solvent
Afterwards remaining residue methyl alcohol, aqueous methanol, acetonitrile or containing water-acetonitrile redissolve, prepare need testing solution；
(2) rhizoma paridis saponin I, II, VI, VII, H, the mixture of Gracillin are taken, reference substance solution is prepared；
(3) need testing solution, reference substance solution are injected separately into Ultra Performance Liquid Chromatography instrument, are detected using external standard method；
Wherein, chromatographic condition is as follows：
Chromatographic column：Octadecylsilane chemically bonded silica is filler,
ELSD detector conditions：55 ± 2 DEG C of drift tube temperature, 40 ± 2psi of nitrogen pressure；
Mobile phase：Acetonitrile (A)-water (B) system, its Gradient program is as follows：
0~1min, (5% or 10%)~(25% or 30%) A；1~(2.5 or 3) min, (25% or 30%)~30%A；
(2.5 or 3)~4min, 30%~(35% or 40%) A；4~(6 or 8) min, (35% or 40%)~40%A；(6 or 8)~
20min, 40%~50%A；20~22min, 50%~70%A；22-23min, 70%~90%A.
6. differentiating method according to claim 5, it is characterised in that：In step (1), alcohols solvent is selected from methyl alcohol, ethanol
Or propyl alcohol, aqueous alcohols solvent is selected from aqueous methanol, hydrous ethanol or aqueous propyl alcohol；In step (2), the solvent of reference substance solution
Selected from methyl alcohol, aqueous methanol, acetonitrile or containing water-acetonitrile.
7. differentiating method according to claim 5, it is characterised in that：In step (3), the column size be 2.1 ×
50mm, the particle diameter of filler is 1.8 μm.
8. differentiating method according to claim 5, it is characterised in that：0.1~0.3mLmin of flow rate of mobile phase-1；Column temperature
40±2℃。
9. differentiating method according to claim 5, it is characterised in that：Eluent gradient is：
0~1min, 10%~30%A；1~3min, 30%~30%A；3~4min, 30%~40%A；4~8min, 40%
~40%A；8~20min, 40%~50%A；20~22min, 50%~70%A；22~23min, 70%~90%A.
10. the method for distinguishing Dali Paris polyphylla and Dulong Paris polyphylla, it is characterised in that：It is detected using liquid chromatography, if detection
To containing chonglou saponin VII, H, 0.010% is below without Gracillin, chonglou saponin I, II or content, then can determine that
It is Dali Paris polyphylla；If detecting containing Gracillin, 0.010% is below without chonglou saponin VII, H, VI or content,
Then can determine that to be Dulong Paris polyphylla.
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| CN108164579A (en) * | 2018-03-23 | 2018-06-15 | 中国食品药品检定研究院 | A kind of method of aerial part extraction separation chonglou saponin H from Paris polyphylla |
| CN109212111A (en) * | 2018-08-23 | 2019-01-15 | 广东东阳光药业有限公司 | Obtain the method and its application of Paris polyphylla extracting solution |
| CN113960233A (en) * | 2021-08-26 | 2022-01-21 | 云南中医药大学 | Method for determining content of 8 types of rhizoma paridis saponins in rhizoma paridis Yunnanensis |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108164579A (en) * | 2018-03-23 | 2018-06-15 | 中国食品药品检定研究院 | A kind of method of aerial part extraction separation chonglou saponin H from Paris polyphylla |
| CN109212111A (en) * | 2018-08-23 | 2019-01-15 | 广东东阳光药业有限公司 | Obtain the method and its application of Paris polyphylla extracting solution |
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| CN113960233A (en) * | 2021-08-26 | 2022-01-21 | 云南中医药大学 | Method for determining content of 8 types of rhizoma paridis saponins in rhizoma paridis Yunnanensis |
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