CN107102015A - The authentication method of paris polyphylla - Google Patents

The authentication method of paris polyphylla Download PDF

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CN107102015A
CN107102015A CN201710167076.7A CN201710167076A CN107102015A CN 107102015 A CN107102015 A CN 107102015A CN 201710167076 A CN201710167076 A CN 201710167076A CN 107102015 A CN107102015 A CN 107102015A
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authentication method
checked
print
paris polyphylla
infrared spectrum
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CN107102015B (en
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刘圆
余孟杰
张绍山
黄钟杰
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Southwest Minzu University
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Southwest Minzu University
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N2021/3595—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using FTIR
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2223/00—Investigating materials by wave or particle radiation
    • G01N2223/05—Investigating materials by wave or particle radiation by diffraction, scatter or reflection
    • G01N2223/056—Investigating materials by wave or particle radiation by diffraction, scatter or reflection diffraction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2223/00—Investigating materials by wave or particle radiation
    • G01N2223/10—Different kinds of radiation or particles
    • G01N2223/101—Different kinds of radiation or particles electromagnetic radiation
    • G01N2223/1016—X-ray

Abstract

The invention provides a kind of single and compound authentication method to paris polyphylla, it is detected using infrared wide spectrum and realized.Using the above method, the certified products of paris polyphylla or adulterant can be subjected to discriminating differentiation, new method is provided for the identification of paris polyphylla by comparing the similarity of standard finger-print and medicinal materials fingerprint to be checked.

Description

The authentication method of paris polyphylla
Technical field
The present invention relates to Chinese medicine quality testing field, and in particular to the authentication method of paris polyphylla.
Background technology
Paris polyphylla is liliaceous plant P.polyphylla Smith var.chinensis (Franch.) Hara. Dry rhizome, version in 2015《Pharmacopoeia of People's Republic of China》Regulation is using chonglou saponin as its criteria of quality evaluation, it and cloud South heavy building is by regular source of the States Pharmacopoeia specifications for Paris polyphylla.Paris polyphylla steroid saponin is the main active of Paris polyphylla, with anti-swollen Knurl, anti-inflammatory, hemostasis, antibacterial, analgesia, calmness, anti-oxidant, immunological regulation, suppression angiogenesis, protection liver and renal effect.
The relevant enterprises such as Yunnan Baiyao group, Sichuan Guangda Pharmaceutical Co., Ltd and TongHua WanTong Pharmacy Co., Ltd It is big to rhizoma paris rhizome resources requirement, wild resource is excessively excavated, the market price rises suddenly and sharply, current rhizoma paris rhizome wild resource is increasingly It is in imminent danger, meanwhile, artificial growth there is no high-quality, stable introduces a collection supply market at present, cause the rhizome of non-pharmacopeia Paris polyphylla plant species Commodity market is full of, the bottleneck of restriction Paris polyphylla pharmaceutical industries sustainable development is had become, the rhizoma paris rhizome of medicinal material market is difficult Accurately differentiated using methods such as conventional character, micro-, physics and chemistry, thin layer discriminatings, therefore, set up the new of paris polyphylla Discrimination method seems especially necessary.
The content of the invention
Present invention aims at provide a kind of single and compound authentication method to paris polyphylla.
Specifically, the invention provides the authentication method of paris polyphylla, it includes following operation：
(1) more than 10 batches paris polyphyllas are taken, crushes, after tabletting, is detected using infrared spectrum, mensuration region 4000- 400cm-1, based on the corresponding wave number of each peak absorption intensity average value and each peak in each batch of sample detection data, obtain red External spectrum standard finger-print；
(2) medicinal material to be checked is taken, is detected using step (1) identical condition, medicinal materials fingerprint to be checked is obtained；
(3) medicinal materials fingerprint to be checked and infrared spectrum standard finger-print are subjected to similarity-rough set, differentiate medicine to be checked The true and false of material；
Wherein, in the infrared spectrum standard finger-print, at least in about 3400cm-1、2940cm-1、1640cm-1、 1411cm-1、1250cm-1、1200-950cm-1、1150cm-1There is absworption peak at place.
Further, in the infrared spectrum standard finger-print, also in about 1020cm-1、920cm-1、860cm-1、 760cm-1、700cm-1There is absworption peak at place.
Infrared spectrogram leads to common wavelengths (λ) or wave number (σ) is abscissa, represents the position of absworption peak, uses light transmittance (T%) or absorbance (A) be ordinate, represent absorption intensity.
In an embodiment of the invention, the infrared spectrum standard finger-print is as shown in Figure 2.
Wherein, before infrared spectrum detection, the mass ratio for choosing paris polyphylla or medicinal material to be checked and KBr is 4：200 are carried out Tabletting.During using said ratio, the light transmittance of most strong absworption peak is below 10, beneficial to detection.
Experiments indicate that, when similarity is more than 0.90, then may determine that medicinal material to be checked is paris polyphylla.
In addition, present invention also offers another of paris polyphylla authentication method, it at least includes using infrared light Spectrum and two kinds of authentication methods of x-ray powder diffraction, each method identify order in no particular order, wherein：Infrared spectrum authentication method：(1) More than 10 batches paris polyphyllas are taken, crushes, after tabletting, is detected using infrared spectrum, mensuration region 4000-400cm-1, with each batch Based on the corresponding wave number of each peak absorption intensity average value and each peak in sample detection data, infrared spectrum standard fingerprint is obtained Collection of illustrative plates；
(2) medicinal material to be checked is taken, is detected using step (1) identical condition, medicinal materials fingerprint to be checked is obtained；
(3) medicinal materials fingerprint to be checked and infrared spectrum standard finger-print are subjected to similarity-rough set, differentiate medicine to be checked The true and false of material；
Wherein, in the infrared spectrum standard finger-print, at least in about 3400cm-1、2940cm-1、1640cm-1、 1411cm-1、1250cm-1、1200-950cm-1、1150cm-1There is absworption peak at place；
X-ray powder diffraction authentication method：More than 10 batches paris polyphyllas are taken, are detected using x-ray powder diffraction, with each Criticize based on each diffraction maximum peak intensity average value and the corresponding 2 θ values of each diffraction maximum in sample detection data, obtain x-ray powder Diffraction standard finger-print；Medicinal material to be checked is taken, is detected using x-ray powder diffraction condition same as before, medicine to be checked is obtained Material finger-print；Medicinal materials fingerprint to be checked and x-ray powder diffraction standard finger-print are subjected to similarity-rough set；
When gained similarity is more than 0.90 in each authentication method, then judge medicinal material to be checked for paris polyphylla.
In an embodiment of the invention, x-ray powder diffraction is carried out using CuK α radiation.
Wherein, 8 °/min of sweep speed.
Wherein, DS=SS=1 ° of slit width, RS=0.3mm.
Wherein, the θ of scanning range 2：5-65°.
Wherein, need to crush before each sample is detected, and cross 100~200 mesh sieves.
In an embodiment of the invention, the finger-print detected using OMNIC software processings infrared spectrum；Using The finger-print of OriginPro software processing x-rays powder diffraction detection.In addition, at the data of x-ray powder pattern fingerprint collection of illustrative plates Reason, can also use the correlation technique in patent 201410105727.6.
In an embodiment of the invention, the x-ray powder diffraction standard finger-print is as shown in Figure 4.
Further, it can also include UPLC authentication methods：
Medicinal material to be checked is taken, is extracted using methanol or ethanol, extract prepares need testing solution, detected through UPLC；Work as chromatogram When at least there is the chonglou saponin VII chromatographic peaks corresponding with H in figure, then judge medicinal material to be checked for paris polyphylla；
Wherein, the UPLC chromatographic conditions are as follows：
Chromatographic column：C18 chromatographic columns, specification 2.1mm × 75mm, 1.7 μm
ELSD conditions：55 DEG C of drift tube temperature, nitrogen pressure 40psi, gain 500, refrigerating mode, 40 DEG C of column temperature, flow velocity 0.2mL·min-1
Mobile phase is acetonitrile-water, and gradient condition is as follows：
In research process of the present invention, using 3 kinds of different chromatographic column BEH C18 chromatographic columns (2.1mm × 50mm, 1.7 μm), HSS C18 chromatographic columns (2.1mm × 75mm, 1.7 μm), BEH C18 chromatographic columns (2.1mm × 100mm, 1.7 μm), result of the test table It is bright：Carry out eluting most preferably using HSS C18 chromatographic columns (2.1mm × 75mm, 1.7 μm), each chromatographic peak retention time is moderate, peak shape It is preferable with separating.
Using above-mentioned complex method, the true and false of paris polyphylla can be more precisely identified, is paris polyphylla Identification provides new method.
The present invention has used two methods of correlation coefficient process, co sinus vector included angle method to calculate similarity, two in an experiment The method of kind is the common method of similarity, and its calculation formula is as follows：
In an embodiment of the invention, for paris polyphylla, similarity is carried out using correlation coefficient process Judgement, its result of determination may be better than co sinus vector included angle method.
The selection of step (1) Chinese medicine batch be should ensure that more than 10 batches, but concrete numerical value is not fixed, as long as meeting system Meter learns meaning.In addition, " paris polyphylla or Yunnan Rhizoma Paridis " described in step (1), refers to through other known into skilled worker Section is accredited as the certified products of paris polyphylla or Yunnan Rhizoma Paridis.
" medicinal material to be checked " described in step (2), can be certified products or adulterant.The present invention is by the above method Certified products and adulterant can be differentiated and distinguished.
Finger-print refers to some complex materials, such as Chinese medicine, the DNA of certain organism or certain tissue or cell, egg After white matter is appropriately processed, using certain analysis means, what is obtained can indicate the chromatogram or spectrogram of its chemical feature. " similarity ", is defined as one in traditional Chinese medicine fingerprint standard by Chinese Pharmacopoeia Commission as a qualitative, quantitative parameter The important evaluation index of item, is applied to the research (National Drug Administration, 2000) of chromatogram class finger-print at present.
In order to ensure the accuracy for detecting data, the inspection of 3 times or more can be carried out to each sample in a conventional manner Survey.
Brief description of the drawings
The infared spectrum of Fig. 1 paris polyphylla rhizomes
The infrared spectrum standard finger-print of Fig. 2 paris polyphylla rhizomes
The X-ray diffraction finger-print of the multiple batches of paris polyphylla rhizomes of Fig. 3
The X-ray diffraction standard finger-print of Fig. 4 paris polyphylla rhizomes
Fig. 5 mixing reference substance UPLC chromatograms, wherein, 1. chonglou saponin VII；3. chonglou saponin H；4. chonglou saponin VI； 5. saikosaponin D；6. chonglou saponin II；7. Gracillin；8. chonglou saponin I
Embodiment
Instrument and material
General analysis XD-6 types x-ray powder diffraction instrument (Beijing Puxi General Instrument Co., Ltd), IR200 Fouriers become Change infrared spectrometer (Thermo Necolet companies of the U.S.)；HX-200 type high speeds medicinal herb grinder (Zhejiang Yongkang small stream bank hardware Medical instrument factory)；DHG-9246 types electric heating constant-temperature blowing drying box (the upper grand experimental facilities Co., Ltd of Nereid).Agate mortar.KBr powder Last (spectroscopic pure) (Thermo companies of the U.S.).
Paris polyphylla and rhizome are taught by Southwest University for Nationalities national medicine research institute Liu Yuan, Sichuan University's West China pharmacy Institute professor Zhang Hao identifies that former plant specimen and medicinal material Sample storage is in Southwest University for Nationalities, HuaXi college of pharmacy, SiChuan University.Sample Information is shown in Table 1.
The paris polyphylla sample Ji Yuan of table 1 and place of production information
The infrared fingerprint of embodiment 1 is identified
Detection method:
Sample treatment：Take after crude drug crushing, cross No. 5 sieves standby.Take the dry 6h of 60 DEG C of each sample powder.Essence takes respectively respectively Sample 4mg (makes the light transmittance of its most strong absworption peak below 10), adds KBr 200mg, and mixed grinding is uniform under infrared lamp, It is put into mould, tabletting, spectral resolution 4cm-1, mensuration region 4000-400cm-1, medium sweep, scanning times 16 times sweep Immediate removal moisture and CO when retouching2Ambient interferences be to ensure the comparativity of collection of illustrative plates.In order to reduce the error that dress sample is caused, each Parallel sweep 3 times after sample grinding is uniform, i.e., pour out dress sample tabletting again after determining every time by sample, to ensure that spectra collection has It is representative, the spectrum mean value that 3 measure are obtained is taken as the absorption spectrum of the sample.
Testing result
Spectral scan, collection data, and the application software of EZ OMNIC 32 carried with infrared spectrometer is carried out to be located Reason, background correction, and carry out baseline correction and automatic smoothing processing.Obtain each sample infrared light collection of illustrative plates figure (see Fig. 1).
The foundation of infrared spectrum standard finger-print
Before similarity analysis is carried out, the average value using the rhizome absorption intensity of 10 portions of paris polyphyllas is standard fingerprint The absorption intensity of collection of illustrative plates, the infared spectrum finally given such as Fig. 2.
From Fig. 1,2 it is observed that wherein 3400cm-1Neighbouring strong and wide absworption peak, the hydroxyl for being attributed to polysaccharide is formed A variety of hydrogen bond O-H keys, the stretching vibration of amino N-H keys absorb；2940cm-1Neighbouring absworption peak is the methylene such as polysaccharide, lipid C-H antisymmetric stretching vibrations；1640cm-1Neighbouring absworption peak be mainly in the materials such as steroidal glycoside, polysaccharide O-H flexural vibrations and Carbonyl C-O stretching vibration absworption peaks；1411cm-1Neighbouring absworption peak is attributed to methylene and becomes angular oscillation；1250cm-1Nearby inhale Receive peak C-OH in carboxylic acid stretching vibration；In 1200-950cm-1Polysaccharide uptake zone shows stronger absworption peak, main To be glycoside, alcohols or glucide C-O stretching vibration absworption peaks；Wherein 1150cm-1Neighbouring absworption peak is polysaccharide and glycoside Hydroxyl O-H flexural vibrations in substance C-O keys eigen vibration and steroid sapogenin.
In addition, in 1020cm-1Neighbouring absworption peak is attributed to straight chain C-C stretching vibration；920、860、760、700cm-1It is attached Near absworption peak is the C-C stretching vibrations in sugared ring and the characteristic absorption vibration of steroid saponin.
Infared spectrum similarity analysis
Using the whole infrared spectrogram of each sample as analysis object, the full spectrum to sample carries out similarity analysis.Instrument Data measured point is as characteristic value, and the difference of the absorption intensity value according to corresponding to each data point position carries out similarity point Analysis.Conventional similarity analysis method has two kinds of " correlation coefficient process " and " co sinus vector included angle method ", and calculation formula is as follows：
Similarity (coefficient correlation and included angle cosine) between each sample is calculated, result of calculation is shown in Table 1.
The infrared Similarity Measure result of table 2
S8 has some gaps with respect to other samples as can be known from Table 2, finds that it is cultivated in soil when being sampled according to seminar In barren, weedy, daily management carelessness environmental condition；Meanwhile, the result that pre-stage test is obtained shows that S8 saponin(e is total Content is substantially lower slightly, and this is probably the reason for causing the change of divergence.When calculating similarity with co sinus vector included angle method, different samples Result of calculation be very close to, and there is no obvious relation with the quality condition of sample, be unfavorable for the comparison of sample room；And use phase When closing Y-factor method Y calculating, its result then has larger difference.Premenstruum (premenstrua) identifies S8 chonglou saponin contents well below States Pharmacopoeia specifications 0.6%, its similarity is 0.92；Remaining infared spectrum is with compareing the similarity of collection of illustrative plates more than 0.95.It can be seen that using related When Y-factor method Y carries out full spectrum similarity analysis to the rhizome infrared finger print atlas of paris polyphylla, its result can reflect exactly Go out the actual mass situation of paris polyphylla rhizome, therefore available for the discriminating and quality evaluation of paris polyphylla medicinal material rhizome.
The compound authentication method of the paris polyphylla of embodiment 2
Although not being found in experiment identify the case not being inconsistent with infrared finger print atlas at present, in order to avoid as far as possible Careless omission in Med Mat Appreciation, it is ensured that the higher accuracy of identification, invention also uses compound authentication method, i.e., by infrared, x-ray The methods such as diffraction are used in combination.
Wherein, infrared authentication method such as embodiment 1.
X-ray powder diffraction methods are as follows：
Detection method：
Sample treatment：Paris polyphylla rhizome sample comminution is taken, 60 DEG C of drying 6h cross 100 mesh sieves standby.Experimental condition： CuK α are radiated；Scan mode：It is qualitative, step-scan；Voltage/current：36kV/20mA；Sweep speed：8°/min；DS=SS= 1 °, RS=0.3mm；Step width：0.02°；The θ of scanning range 2：5-65°.
Testing result：
1st, total Test data handle (smooth) at 35 points with OriginPro 8.5, obtain powder diffraction spectrum, as a result as schemed 3。
2nd, the foundation of X diffraction standards finger-print
Before similarity analysis is carried out, referred to using the average value of 10 parts of paris polyphylla rhizome peak intensities as X diffraction standards The peak intensity of line collection of illustrative plates, the X diffraction standards finger-print such as Fig. 4 finally given.
3rd, X diffracting spectrums similarity analysis
Conventional similarity analysis method has two kinds of " correlation coefficient process " and " co sinus vector included angle method ", and calculation formula is such as Under：
Using similarity (coefficient correlation and included angle cosine) between form calculating each sample.
The Similarity Measure result of table 3
Above-described embodiment calculates the peak for obtaining X diffraction standard finger-prints according to the matching result at peak using mean value method Value；Interplanar distance d diffraction relative intensities I/I0Represent diffraction peak；Using the θ of angle 2 as abscissa, diffraction peak (d/I/I0) it is vertical Coordinate, forms paris polyphylla rhizome X diffraction standard finger-prints, and carried out the comparison of similarity.More than vector angle When string method calculates similarity, S8 is low compared with other samples with compareing similarity, illustrates that the quality condition with sample has certain relation, but It is that data result of calculation is extremely approached, gap is not obvious, is unfavorable for the comparison of sample room；And when being calculated with correlation coefficient process, its As a result then there is larger difference.Premenstruum (premenstrua) identifies S8 chonglou saponins content well below States Pharmacopoeia specifications 0.6%, and its similarity exists 0.9987, this analysis result is consistent with infrared spectrum identification result；The similarity of remaining X diffracting spectrum and standard finger-print More than 0.9992.0.9900 is all higher than according to the visible similarity of result of the comparison, phase very high between embodying Like property, the relative stability of the different Paris polyphylla rhizome quality of Sichuan is further illustrated, is built while also illustrate that Vertical X diffraction standard finger-prints are reliable.
The compound authentication method of the paris polyphylla of embodiment 3
Except that, by addition to the infrared method that identification is combined with x-ray powder diffraction, can also be added in the present invention in embodiment 2 UPLC。
Infrared and x-ray powder diffraction authentication method such as embodiment 2.
UPLC authentication methods are as follows:
Chromatographic condition
ACQUITYHSS C18 chromatographic columns (2.1mm × 75mm, 1.7 μm)；ELSD detectors drift tube temperature 55 DEG C, nitrogen pressure 40psi, gain 500, refrigerating mode；40 DEG C of column temperature；Flow velocity 0.2mLmin-1；Sample size：2μL；Mobile phase For acetonitrile (A)-water (B), gradient elution program is shown in Table 4.Mixing reference substance figure is shown in Fig. 5.
The gradient elution program of table 4
The preparation of need testing solution
Dry paris polyphylla rhizome is crushed, 40 mesh sieves are crossed, precision weighs rhizoma paris rhizome powder 0.500g, is placed in In 100mL conical flasks, by solid-liquid ratio 1：100(g：ML 50mL methanol) is added, is shaken up, is stood after 4h, ultrasonic 30min, filtering takes Filtrate is placed in flask, repeats aforesaid operations 1 time.Merging filtrate, 45 DEG C of vacuum rotary steams are to dry, after residue is dissolved with chromatogram methanol, Constant volume is standby through 0.22 μm of filtering with microporous membrane in 10mL volumetric flasks.
The preparation of reference substance solution
The preparation of external standard method reference substance solution
Precision weighs each chonglou saponin reference substance respectively, is dissolved with chromatogram methanol, reference substance concentration in each reference substance solution Respectively 0.206 (VII), 0.210 (H), 0.206 (VI), 0.208 (II), 0.204 (Gracillin), 0.202 (I) mg·mL-1, each Paris polyphylla reference substance solution is produced, it is standby through 0.22 μm of filtering with microporous membrane.
The preparation of internal standard method reference substance solution
Precision weighs each chonglou saponin reference substance respectively, is dissolved with chromatogram methanol, reference substance concentration in each reference substance solution Respectively 0.136 (VII), 0.113 (H), 0.104 (VI), 0.121 (II), 0.106 (Gracillin), 0.103 (I) mg·mL-1, each Paris polyphylla reference substance solution is produced, it is standby through 0.22 μm of filtering with microporous membrane.Precision weighs saikosaponin D, uses color Methanol dissolving is composed, reference substance concentration is 0.250 (saikosaponin D) mgmL-1, produce saikosaponin D reference substance solution.
Methodological study
Linear relationship is investigated
External standard method：Precision draws reference substance solution and is diluted to 2mL in right amount, is configured to the mixing control of different extension rates Product solution, the μ L of sample introduction 2, is determined by chromatographic condition.Using 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 5.
The equation of linear regression of the external standard method reference substance of table 5
Internal standard method：Precision draws each chonglou saponin reference substance solution 0.5,1,1.5,2.5,3mL, accurate respectively to add internal standard Thing (saikosaponin D) solution 1mL, is settled to 4mL, produces Series Measurement reference substance solution, the μ L of sample introduction 2, by foregoing chromatostrip Part is determined.Peak area ratio using internal standard compound (saikosaponin D) and reference substance chonglou saponin is ordinate (Y), their sample introduction Amount ratio is abscissa (X), draws internal standard method standard curve, the results are shown in Table 6.
The equation of linear regression of the internal standard method reference substance of table 6
Precision test
Reference substance solution is taken, precision is drawn, continuous sample introduction 5 times determines record, obtains chonglou saponin VII, H, VI, II, fibre Thin Dioscin, the RSD of I peak areas are respectively 1.14%, 1.36%, 1.28%, 1.77%, 1.81%, 1.55%, as a result ＜ 3%, show that instrument precision is good.
Replica test
Sample S3, parallel 5 parts of rhizome need testing solutions of preparation and the serial need testing solution of internal standard method are taken, by foregoing chromatostrip Part is determined, and records each chonglou saponin peak area, calculates RSD, as a result equal ＜ 3%, shows that method is reproducible.
Stability test
Same need testing solution (S3) is taken, by foregoing chromatographic condition, is determined respectively at 0,2,4,8,12,24h sample introductions, record Each chonglou saponin peak area, calculates RSD, as a result equal ＜ 3%, shows that Paris polyphylla need testing solution stability is good in 24h.
It is loaded recovery test
Take 5 parts of the Paris polyphylla medicinal material sample (S3) with a collection of known content, respectively plus precision weighing add chonglou saponin VII, H, VI, II, Gracillin, I reference substances, sample preparation, constant volume are determined, and calculate the rate of recovery.Chonglou saponin is calculated by external standard method VII, H, VI, II, Gracillin, I mean sample recovery rates are respectively：99.91%th, 99.27%, 98.44%, 101.33%th, 98.78%, 99.01%, RSD is 1.76%, 1.65%, 1.44%, 1.98%, 1.52%, 1.73%；By interior Mark method is calculated, and chonglou saponin VII, H, VI, II, Gracillin, I mean sample recovery rates are respectively：97.21%th, 98.56%th, 100.12%, 100.15%, 99.34%, 98.78%, RSD be 0.92%, 1.77%, 1.81%, 1.65%, 1.34%th, 1.80%.
Testing result
6 contents of steroid saponin (%) in 7 10 parts of sample rhizomes of table
In 10 batches of samples of detection, the chonglou saponin VII of one of pharmacopeia Testing index is detected in all samples, and Chonglou saponin H is also detected in all samples.Remaining saponin constituent is not all present in each sample, therefore, this hair It is bright to think, can be simultaneously using chonglou saponin VII and H as the control composition of the identification paris polyphylla true and false, if with this UPLC side Method combines infrared and x-ray powder diffraction, it will be more beneficial for improving the accuracy of paris polyphylla authenticity.

Claims (10)

1. the authentication method of paris polyphylla, it is characterised in that：It includes following operation：
(1) more than 10 batches paris polyphyllas are taken, crushes, after tabletting, is detected using infrared spectrum, mensuration region 4000-400cm-1, Based on the corresponding wave number of each peak absorption intensity average value and each peak in each batch of sample detection data, infrared spectrum mark is obtained Quasi- finger-print；
(2) medicinal material to be checked is taken, is detected using step (1) identical condition, medicinal materials fingerprint to be checked is obtained；
(3) medicinal materials fingerprint to be checked and infrared spectrum standard finger-print are subjected to similarity-rough set, differentiate medicinal material to be checked The true and false；
Wherein, in the infrared spectrum standard finger-print, at least in about 3400cm-1、2940cm-1、1640cm-1、1411cm-1、 1250cm-1、1200-950cm-1、1150cm-1There is absworption peak at place.
2. authentication method according to claim 1, it is characterised in that：In the infrared spectrum standard finger-print, also exist About 1020cm-1、920cm-1、860cm-1、760cm-1、700cm-1There is absworption peak at place；Further, the infrared spectrum standard refers to Line collection of illustrative plates is as shown in Figure 2.
3. authentication method according to claim 1, it is characterised in that：Infrared spectrum detection before, choose paris polyphylla or The mass ratio of medicinal material and KBr to be checked is 4：200 carry out tabletting.
4. authentication method according to claim 1, it is characterised in that：When similarity is more than 0.90, then judge to be checked Medicinal material is paris polyphylla.
5. the authentication method of paris polyphylla, it is characterised in that：It at least includes using two kinds of infrared spectrum and x-ray powder diffraction Authentication method, each method identifies order in no particular order, wherein：
Infrared spectrum authentication method：As described in Claims 1 to 4 any one；
X-ray powder diffraction authentication method：More than 10 batches paris polyphyllas are taken, are detected using x-ray powder diffraction, with each lot sample Product examine is surveyed based on each diffraction maximum peak intensity average value in data and the corresponding 2 θ values of each diffraction maximum, obtains x-ray powder diffraction Standard finger-print；Medicinal material to be checked is taken, is detected using x-ray powder diffraction condition same as before, medicinal material to be checked is obtained and refers to Line collection of illustrative plates；Medicinal materials fingerprint to be checked and x-ray powder diffraction standard finger-print are subjected to similarity-rough set；
When gained similarity is more than 0.90 in each authentication method, then judge medicinal material to be checked for paris polyphylla.
6. authentication method according to claim 5, it is characterised in that：X-ray powder diffraction is carried out using CuK α radiation.
7. authentication method according to claim 6, it is characterised in that：Sweep speed 8 °/min, slit width DS=SS= 1 °, RS=0.3mm, the θ of scanning range 2：5-65°.
8. authentication method according to claim 5, it is characterised in that：Detected using OMNIC software processings infrared spectrum Finger-print；The finger-print detected using OriginPro software processing x-rays powder diffraction.
9. authentication method according to claim 5, it is characterised in that：The x-ray powder diffraction standard finger-print such as Fig. 4 It is shown.
10. authentication method according to claim 5, it is characterised in that：It also includes UPLC authentication methods：
Medicinal material to be checked is taken, is extracted using methanol or ethanol, extract prepares need testing solution, detected through UPLC；When in chromatogram When at least there is the chonglou saponin VII chromatographic peaks corresponding with H, then judge medicinal material to be checked for paris polyphylla；
Wherein, the UPLC chromatographic conditions are as follows：
Chromatographic column：C18 chromatographic columns, specification 2.1mm × 75mm, 1.7 μm
ELSD conditions：55 DEG C of drift tube temperature, nitrogen pressure 40psi, gain 500, refrigerating mode, 40 DEG C of column temperature, flow velocity 0.2mL·min-1
Mobile phase is acetonitrile-water, and gradient condition is as follows：
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108490007A (en) * 2018-06-04 2018-09-04 西南民族大学 The method that subdigitate wormwood herb and subdigitate wormwood herb charcoal are distinguished using X-ray diffraction method
CN108760670A (en) * 2018-06-04 2018-11-06 西南民族大学 Using the method for infrared differentiation subdigitate wormwood herb and subdigitate wormwood herb charcoal
CN109055603A (en) * 2018-10-11 2018-12-21 湖北省农业科学院中药材研究所 A method of building paris plant finger-print is marked using CDDP
CN110243781A (en) * 2019-05-20 2019-09-17 广东产品质量监督检验研究院 The detection method of leather and fur products
CN111337448A (en) * 2020-03-26 2020-06-26 杭州老爸标准技术集团有限公司 Method for identifying cosmetics based on infrared spectrum technology

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103487395A (en) * 2013-09-05 2014-01-01 浙江大学 Quick multi-index detection method for Paris polyphylla medicinal materials
CN103983603A (en) * 2014-05-29 2014-08-13 南京中医药大学 Establishment method of fourier transform infrared spectrum fingerprint of mineral medicine limonitum, standard fingerprint and application thereof
CN106290221A (en) * 2016-07-27 2017-01-04 广东药科大学 A kind of Chinese traditional medicine mirabilite Quality evaluation based on FT IR fingerprint chromatogram method
CN106353417A (en) * 2016-08-14 2017-01-25 西南民族大学 UPLC (ultra-performance liquid chromatography) detection method of multiple steroid saponins in Yunnan rhizoma paridis or its polygerm strain

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103487395A (en) * 2013-09-05 2014-01-01 浙江大学 Quick multi-index detection method for Paris polyphylla medicinal materials
CN103983603A (en) * 2014-05-29 2014-08-13 南京中医药大学 Establishment method of fourier transform infrared spectrum fingerprint of mineral medicine limonitum, standard fingerprint and application thereof
CN106290221A (en) * 2016-07-27 2017-01-04 广东药科大学 A kind of Chinese traditional medicine mirabilite Quality evaluation based on FT IR fingerprint chromatogram method
CN106353417A (en) * 2016-08-14 2017-01-25 西南民族大学 UPLC (ultra-performance liquid chromatography) detection method of multiple steroid saponins in Yunnan rhizoma paridis or its polygerm strain

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
何立巍等: "中药石膏炮制品的X射线衍射分析及指纹图谱的建立", 《天津中医药》 *
张绍山等: "UPLC 法测定云南省不同地区云南重楼及多芽品系中7种甾体皂苷量及其指纹图谱建立", 《中草药》 *
梁玉勇等: "HPLC 测定贵州不同产地的七叶一枝花中9中甾体皂苷的含量", 《中国中药杂志》 *
高夏红: "中草药XRD指纹图谱方法学研究及中药XRD指纹图谱数据库建立", 《中国优秀博硕士学位论文全文数据库（硕士）医药卫生科技辑》 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108490007A (en) * 2018-06-04 2018-09-04 西南民族大学 The method that subdigitate wormwood herb and subdigitate wormwood herb charcoal are distinguished using X-ray diffraction method
CN108760670A (en) * 2018-06-04 2018-11-06 西南民族大学 Using the method for infrared differentiation subdigitate wormwood herb and subdigitate wormwood herb charcoal
CN108490007B (en) * 2018-06-04 2021-02-26 西南民族大学 Method for distinguishing artemisia rupestris L and artemisia rupestris L charcoal by adopting X-ray diffraction method
CN109055603A (en) * 2018-10-11 2018-12-21 湖北省农业科学院中药材研究所 A method of building paris plant finger-print is marked using CDDP
CN110243781A (en) * 2019-05-20 2019-09-17 广东产品质量监督检验研究院 The detection method of leather and fur products
CN111337448A (en) * 2020-03-26 2020-06-26 杭州老爸标准技术集团有限公司 Method for identifying cosmetics based on infrared spectrum technology
CN111337448B (en) * 2020-03-26 2023-05-12 杭州老爸评测科技股份有限公司 Method for identifying cosmetics based on infrared spectrum technology

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