CN109374764A - Eight taste dragons of one kind bore particle HPLC finger-print and Contents of Main Components measuring method - Google Patents
Eight taste dragons of one kind bore particle HPLC finger-print and Contents of Main Components measuring method Download PDFInfo
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- CN109374764A CN109374764A CN201811226190.3A CN201811226190A CN109374764A CN 109374764 A CN109374764 A CN 109374764A CN 201811226190 A CN201811226190 A CN 201811226190A CN 109374764 A CN109374764 A CN 109374764A
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- 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
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- 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
Abstract
The present invention provides a kind of method that can be used for eight taste dragons simultaneously and bore particle main component HPLC Qualitative fingerprint and quantitative analysis.The method uses 70% methanol ultrasound assisted extraction, using octadecylsilane chemically bonded silica as stationary phase, -0.1% phosphate aqueous solution of acetonitrile is mobile phase, only can measure the qualitative finger-print of feature of said preparation and the content of 7 kinds of main actives simultaneously with a chromatographic condition.It is proved through scientific experiment, this method is easy to operate, quick, and repeatability and stability are good, and Fingerprints are strong, and assay result is accurate, can be used for the quality control that eight taste dragons bore grain products.
Description
Technical field
The present invention relates to one kind can be used for the method that eight taste dragons bore particle main component quantification and qualification simultaneously, more really
Say it is a kind of eight taste dragons brill particle HPLC Qualitative fingerprint and 7 kinds of primary efficacy Related Component gallic acids, former catechu with cutting
Detection method while acid, cucoline, rhodioside, catechin, aurantiamarin and psoralen are quantitative, belongs to Chinese traditional compound medicine
Analysis detection field.
Background technique
It is on the basis of excavating the conventional medicament for summarizing strong medical treatment rheumatoid arthritis and " strong that eight taste dragons, which bore particle,
The intension of doctor's poison opinion ", the strong patent medicine researched and developed according to the clinical proved recipe of strong medical knowledge opinion compatibility.The party mainly by toddalia, wammel,
Radix zanthoxyli, caulis sinomenii, Caulis Spatholobi, champion bauhinia stem, alangium and eight taste medicine of radix fici simplicissimae composition, all have anti-inflammatory, analgesia, wind resistance
It is the effect of wet, immunosupress, significant to treatment rheumatoid arthritis.Champion bauhinia stem, Caulis Spatholobi in side etc. are usually used in treating
Rheumatic immunological disease, champion bauhinia stem have stronger antalgic and inflammation relieving activity, are the strong wet immunological disease training common medicine of the style of work, Caulis Spatholobi tool
There are blood circulation promoting and enriching, dredging collateral, dispelling wind and eliminating dampness effect；Caulis sinomenii has effects that wind-dispelling dissipates the stasis of blood, dampness removing analgesic, and radix fici simplicissimae can be good for
Spleen QI invigorating, removing dampness and relaxing myospasm, promoting qi circulation and relieving pain.Modern pharmacological studies have shown that the biology total alkali of toddalia, ethanol extract and water mention
Object has analgesic and anti-inflammatory effects；Radix zanthoxyli has swelling and pain relieving effect, there is antifatigue, anti-rhodioside in sargentodoxa cuneata (wammel)
The multiple pharmacological effects such as inflammation, anti-aging, immunological regulation are one of active constituents of wammel；Cucoline has anti-inflammatory, analgesia, resists
The pharmacological actions such as rheumatism；Psoralen is often used as the quality control index of radix fici simplicissimae medicinal material；And aurantiamarin has anti-inflammatory, tune
The pharmacological activity waited is immunized in section；Nitidine Chloride is significant in the effect of anti-inflammatory aspect；Gallic acid and protocatechuic acid all have bright
Aobvious anti-oxidant, inhibition platelet aggregation；Catechin has the function of certain promotion hematopoietic cell proliferation, is Caulis Spatholobi
The main matter basis of replenishing and activating blood.And gallic acid has antibacterial, anti-oxidant and antitumor pharmacological action.It is above it is all kinds of at
Divide closely related with said preparation effect.Up to the present, there has been no only can be measured and can be embodied simultaneously with a chromatographic system
Eight taste dragons bore the finger-print of particle medicinal material characteristic information and measure the measuring method of main feature component content in preparation simultaneously
Report.
Summary of the invention
Solution of the invention, which is to provide a kind of can measure simultaneously, can embody eight taste dragons brill particle main Chinese medicinal materials information
Characteristic fingerprint pattern and the measuring method for measuring the relevant main feature component content of 7 kinds of effects in preparation simultaneously, the method use
70% methanol ultrasound assisted extraction, using octadecylsilane chemically bonded silica as stationary phase, -0.1% phosphate aqueous solution of acetonitrile is stream
Dynamic phase can only measure the qualitative finger-print of feature and 7 kinds of main actives of said preparation simultaneously with a chromatographic condition
Content.It is proved through scientific experiment, this method is easy to operate, quick, and repeatability and stability are good, and Fingerprints are strong, contain
Amount measurement result is accurate, can be used for the quality control that eight taste dragons bore grain products.
Eight taste dragons of the invention bore particle characteristic finger-print and the detection method of 7 kinds of effect Related Component assays includes
Following steps:
1. the preparation of reference substance solution: precision weigh gallic acid, protocatechuic acid, cucoline, rhodioside, catechin,
Aurantiamarin and psoralen reference substance are appropriate, and methanol is added to dissolve, and the reference substance stock solution of debita spissitudo is respectively prepared.It separately takes above-mentioned
Appropriate solution is placed in same measuring bottle, adds methanol dilution, and gallic acid containing a certain concentration, protocatechuic acid, cucoline, red is made
Red-spotted stonecrop glycosides, catechin, the mixed reference substance solution of aurantiamarin and psoralen, are placed in freezer storage, spare.
2. the preparation of test solution: precision weigh eight taste dragons bore particle it is appropriate, set in stuffed conical flask, precision be added
70% methanol is appropriate, weighed weight, and ultrasound lets cool room temperature, weighs again, the weight of less loss is supplied with 70% methanol, is shaken up,
Filtering, subsequent filtrate centrifugation, takes supernatant as test solution.
3. chromatographic condition: using octadecylsilane chemically bonded silica as stationary phase, -0.1% phosphate aqueous solution of acetonitrile is flowing
Phase, gradient elution；Column temperature is 25~30 DEG C；Flow velocity is 1ml/min；Detector is UV detector, and Detection wavelength is respectively
270nm and 245nm.
4. measuring method: accurate respectively to draw gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, orange peel
The mixed reference substance solution and test solution of glycosides and psoralen are each appropriate, inject high performance liquid chromatography by above-mentioned chromatographic condition
Instrument, measures corresponding peak area, calculate to get.
After study, the eight taste dragons that the present invention optimizes bore particle characteristic finger-print and 7 kinds of effect Related Components containing measurement
Fixed detection method is implemented according to the following steps:
1. the preparation of reference substance solution: precision weigh gallic acid, protocatechuic acid, cucoline, rhodioside, catechin,
Aurantiamarin and psoralen reference substance are appropriate, and methanol is added to dissolve, and debita spissitudo reference substance stock solution is respectively prepared.It separately takes above-mentioned molten
Appropriate liquid is placed in same measuring bottle, adds methanol dilution, is made containing 0.05~0.10mg/mL of gallic acid, containing protocatechuic acid and benefit
Bone fat element concentration is 0.005~0.01mg/mL, is 0.1~0.3mg/mL, containing orange containing cucoline, rhodioside, catechin concentration
Skin glycosides concentration is the mixed reference substance solution of 0.01~0.05mg/mL, is placed in freezer storage, spare.
2. the preparation of test solution: take eight taste dragons to bore particle 3g, it is accurately weighed, it sets in stuffed conical flask, precision is added
70% methanol of 50mL, weighed weight are ultrasonically treated (power 200W, frequency 40kHz) 20min, let cool, then weighed weight, use
70% methanol supplies the weight of less loss, shakes up, filtration, and subsequent filtrate centrifugation takes supernatant to get test solution；
3. chromatographic condition and chromatography employment and suitability test (E & ST): with Phenomenex Gemini C18 (250mm × 4.6mm i.d.,
5 μm) chromatographic column be stationary phase, -0.1% phosphoric acid water (B) of acetonitrile (A) be mobile phase, the percent by volume condition of gradient elution
It is: 0~10min, 6%A:10~20min, 12%A；20~30min, 19%A；30~40min, 27%A；40~50min,
35%A；50~60min, 45%A；Column temperature is 25~30 DEG C；Flow velocity is 1ml/min；Detector is UV detector, detects wave
A length of 0~50min:270nm, 50~60min:245nm；10 μ L of sample volume；Chromatographic column column effect is not less than in terms of cucoline
30000。
4. measuring method: accurate respectively to draw above-mentioned gallic acid containing a certain concentration, protocatechuic acid, cucoline, root of kirilow rhodiola
Glycosides, catechin, aurantiamarin and psoralen each 10 μ l of mixed reference substance solution and test solution, by above-mentioned chromatographic condition
Inject high performance liquid chromatograph, measure corresponding peak area, calculate to get.
Compared with prior art, the present invention its major advantage and having the active effect that
(1) detection method of the invention can only be measured with a chromatographic condition simultaneously by using wavelength conversion technology
Gallic acid in the characteristic fingerprint pattern and preparation of said preparation, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and
The content of 7 kinds of effect Related Components of psoralen.
(2) detection method of the invention, easy, quick, easily operated, not cumbersome operating process.
(3) detection method of the invention has easy, quick, stable, and repeatability and favorable reproducibility, accuracy are high, is easy to
The characteristics of grasping and operating.
Detection method of the invention is the practicable method groped by many experiments, has repeatability
With preferable stability, according to the present invention disclosed technology contents, those skilled in the art will be clear that of the invention other
Embodiment, but as long as using detection method of the present invention, within that scope of the present invention.
Detailed description of the invention:
7 kinds of mixed reference substance solution HPLC-UV maps such as 1 cucoline of attached drawing
2 eight taste dragon of attached drawing bores 7 kinds of component contents in particle test solution and measures HPLC-UV map
3 eight taste dragon of attached drawing bores particle HPLC finger-print stacking chart
4 eight taste dragon of attached drawing bores particle HPLC finger-print common pattern
Specific embodiment:
Below in conjunction with embodiment, the invention will be further described, and the embodiment of the present invention is only as an example, be intended to
The bright present invention, is not construed as limiting the invention.
1 eight taste dragon of embodiment bores the assay of 7 kinds of main components in particle
1 instrument and reagent
1.1 instrument
Agilent-1260 high performance liquid chromatograph；Diode array UV detector；Ultrasonic cleaning machine (KQ5200, elder brother
Ultrasonic instrument Co., Ltd of mountain city)；Centrifuge (TGL-16G, Anting Scientific Instrument Factory, Shanghai)；Electronic balance (ten a ten thousandths,
SQP, Sai Duolisi scientific instrument (Beijing) Co., Ltd)；Mi Sibo ultrapure water machine etc..
1.2 reagent
Reference substance gallic acid (Gallic acid) (110831-201605, content 90.8%), protocatechuic acid
(Protocatechuic acid) (110809-201205, content 99.9%), cucoline (Sinomenine) (110774-
200507, content 99.1%), rhodioside (Salidroside) (110818-2015007, content 99.4%), catechin
((+)-Catechin) (110877-201604, content 99.2%), (110721-201617 contains aurantiamarin (Hesperidin)
Amount is 96.7%) and Psoralen cured plain (Psoralen) (110739-201617, content 99.1%) is purchased from Chinese food drug assay
Research institute；Acetonitrile (chromatographically pure；Fisher company)；Methanol, phosphoric acid (analyzing pure, Sinopharm Chemical Reagent Co., Ltd.)；Water
For ultrapure water.
10 batches of eight taste dragons bore particle, and (every bag of 10g/, every bag is equivalent to 35 grams of crude drug, and experiment is shown in Table with each sample lot number
4) it, is provided by Guangxi University of TCM Pharmaceutical Factory；Experiment crude drug alangium used, wammel, toddalia, Caulis Spatholobi,
Radix zanthoxyli, champion bauhinia stem, radix fici simplicissimae, caulis sinomenii are purchased from Guangxi Yulin City of Guangxi Zhuang Autonomous Xiang Sheng prepared slices of Chinese crude drugs Co., Ltd,
It is identified through Guangxi University of TCM Pharmaceutical Factory quality assurance portion east Luo Yu associate professor of pharmacy, toddalia, wammel, Radix zanthoxyli etc. 8
Kind medicinal material meets the related request of " Guangxi precious jade quality of medicinal material standard ".
2 methods and result
2.1 chromatographic conditions and system suitability
With Phenomenex Gemini C18 (250mm × 4.6mm i.d., 5 μm) chromatographic column for stationary phase, acetonitrile (A)-
0.1% phosphoric acid water (B) is mobile phase, and the percent by volume condition of gradient elution is: 0~10min, 6%A；10~20min,
12%A 20~30min, 19%A30~40min, 27%A；40~50min, 35%A；50~60min, 45%A；Column temperature is 25
~30 DEG C；Flow velocity is 1ml/min；Detector is UV detector, and Detection wavelength is 0~50min:270nm, 50~60min:
245nm；10 μ L of sample volume；Chromatographic column column effect is not less than 30000 in terms of cucoline.Under this chromatographic condition, in prescription it is other at
Divide noiseless to measuring.Eight taste dragons bore the mixed reference substance solution and eight taste dragons brill particle test sample of 7 kinds of main components in particle
The HPLC-UV map of solution is shown in attached drawing 1 and attached drawing 2 respectively.
The preparation of 2.2 reference substances and test solution
2.2.1 the preparation of mixed reference substance solution
Precision weighs gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and psoralen control
Appropriate product add methanol dissolution that reference substance stock solution is made.It separately takes above-mentioned solution appropriate, is placed in same measuring bottle, adds methanol dilution,
It is made and is respectively containing gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and psoralen concentration
0.0696mg/mL、0.0087mg/mL、0.2046mg/mL、0.2564mg/mL、0.1004mg/mL、0.0243mg/mL、
The mixed reference substance solution of 0.0071mg/mL is placed in freezer storage, spare.
2.2.2 the preparation of test solution
Precision weighs eight taste dragons and bores particle 3g, sets in stuffed conical flask, precision addition 70% methanol of 50mL, weighed weight,
Ultrasonic 20min (power 200W, frequency 40kHz), lets cool room temperature, weighs again, the weight of less loss is supplied with 70% methanol, is shaken
Even, filtering, subsequent filtrate centrifugation takes supernatant as test solution.
2.3 measuring method
It is accurate respectively to draw above-mentioned gallic acid containing a certain concentration, protocatechuic acid, cucoline, rhodioside, catechin, orange
Each 10 μ l of the mixed reference substance solution and test solution of skin glycosides and psoralen injects efficient liquid phase by above-mentioned chromatographic condition
Chromatograph, measures corresponding peak area, calculate to get.
2.4 methodological study
2.4.1 system suitability
Reference substance solution, sample solution are taken respectively, and by chromatographic condition under " 2.1 " item, sample introduction is analyzed, is examined with diode array
It surveys device to be detected, as the result is shown gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and psoralea corylifolia
The chromatographic peak retention time and UV spectrum and chromatographic peak retention time corresponding in sample of this plain 7 ingredients are consistent with UV spectrum, respectively
Peak separating degree is preferable, and tailing factor is between 0.95~1.05, and other compositions are to the measurement of this 7 ingredients without dry in sample
It disturbs, chromatographic column theoretical cam curve is not less than 30000 in terms of cucoline, and the results are shown in attached figure 1 and attached drawing 2.
2.4.2 the investigation of linear relationship
The mixed reference substance solution under " 2.2.1 " item is taken, using autosampler, precision takes 4,6,8,10,12,14 respectively
The analysis of μ L injecting chromatograph records the peak area of corresponding chromatographic peak by the chromatographic condition sample introduction measurement under " 2.1 " item, respectively with
Peak area Y is ordinate, and sample volume (μ g) X is abscissa, acquires equation of linear regression and linearly dependent coefficient (r), as a result sees
Table 1.
1 regression equation of table and the range of linearity
2.4.2 precision test
The mixed reference substance solution under " 2.2.1 " item is taken, measures, continuous sample introduction 6 times, is surveyed respectively by " 2.1 " chromatographic condition
Gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and psoralen peak area RSD be respectively
1.3%, 1.3%, 0.77%, 0.87%, 1.1%, 0.94% and 1.2%, display instrument precision is good.
2.4.3 stability test
It takes eight taste dragons to bore particle (lot number 20170501), by legal system available test sample solution below " 2.2.2 " item, presses " 2.1 "
Chromatographic condition measurement successively measures gallic acid, protocatechuic acid, cucoline, red scape respectively in 0h, 2h, 4h, 8h, 12h, for 24 hours
Its glycosides, catechin, aurantiamarin and psoralen peak area RSD be respectively 1.2%, 1.1%, 1.4%, 1.8%, 1.0%,
1.0% and 1.2%, display sample solution is stablized interior for 24 hours.
2.4.4 repetitive test
Precision weighs eight taste dragons and bores 6 parts, each 3.0g of particle (lot number 20170501), prepares 6 parts by method below " 2.2.2 " item
Test solution is measured, as a result gallic acid, protocatechuic acid, cucoline, rhodioside, catechu by chromatographic condition under " 2.1 " item
Element, aurantiamarin and psoralen peak average content be respectively as follows: 0.6959mg/g, 0.1364mg/g, 3.5620mg/g,
3.6991mg/g, 1.2136mg/g, 0.3234mg/g, 0.07650mg/g, RSD is respectively 0.71%, 1.9%, 2.0%,
1.5%, 2.0%, 1.5% and 1.5%, show that the repeatability of this method is good.
2.4.5 sample recovery rate is tested
Eight taste dragons are taken to bore particle (lot number 20170501) (gallic acid, protocatechuic acid, cucoline, rhodioside, catechu
Element, aurantiamarin and psoralen average content be respectively 0.6959,0.1364,3.5620,3.6991,1.2136,0.3234,
0.07650mg/g) about 1.5g, it is totally 6 parts, accurately weighed, it is accurate respectively that gallic acid, protocatechuic acid, cucoline, root of kirilow rhodiola is added
Glycosides, catechin, aurantiamarin and each 1.0500mg, 0.2050mg of psoralen reference substance, 5.5000mg, 5.6000mg,
1.8200mg, 0.4800mg and 0.1200mg.By legal system available test sample solution below " 2.2.2 " item, by chromatostrip under " 2.1 " item
Part measurement, measures each peak area, the sample recovery rate for calculating each ingredient is respectively 103.6%, 102.1%, 102.5%,
99.6%, 103.2%, 99.2% and 98.5%, RSD be respectively 1.1%, 1.0%, 1.4%, 1.7%, 1.4%, 0.92%,
And 1.3%, show that method accuracy is good, is shown in Table 2.
The recovery test result (n=6) of 27 ingredients of table
The measurement of 2.5 samples
10 batches of eight taste dragons are taken to bore each 2 parts of particle, respectively about 3g, accurately weighed, molten by legal system available test product below " 2.2.2 " item
Liquid is measured by chromatographic condition under " 2.1 " item, according to the peak area measured, is calculated the average content of each ingredient in sample, is as a result seen
Table 3.
3 sample size measurement result (n=2) (mg/g) of table
2 eight taste dragon of embodiment bores the research of particle HPLC finger-print and measurement
1 instrument and reagent: with embodiment one.
2 methods and result: with embodiment one.
2.1 chromatographic conditions and system suitability: with embodiment one.
The preparation of 2.2 reference substances and test solution: with embodiment one.
2.3 measuring methods: with embodiment one.
2.4 8 taste dragons bore particle HPLC fingerprint spectrum method and investigate and measure
2.4.1 stability test
It takes eight taste dragons to bore particle (lot number 20170501), prepares test solution by the method under " 2.2.2 " item,
Under " 2.1 " item chromatographic condition, respectively 0,2,4,8,12, measure for 24 hours, with the retention time and chromatographic peak of No. 6 peaks (cucoline)
Area is reference, be calculated except the relative retention time RSD referring to remaining 11 shared peak in addition to peak cucoline 0.33%~
Between 1.8%, between 0.88%~1.7%, experimental result is shown relative peak area RSD, and it is molten that eight taste dragons bore particle test sample
Liquid is stablized interior for 24 hours, meets the related request and regulation of determining fingerprint pattern.
2.4.2 precision test
It takes eight taste dragons to bore particle (lot number 20170501), prepares test solution by the method under " 2.2.2 " item,
Under " 2.1 " item chromatographic condition, continuous sample introduction 6 times, HPLC chromatogram is measured, with the retention time and chromatography of No. 6 peaks (cucoline)
Peak area is reference, and the relative retention time RSD at remaining 11 shared peak in addition to cucoline is calculated 0.40%~1.6%
Between, relative peak area RSD shows that instrument precision is good between 0.62%~1.8%, and meets determining fingerprint pattern
Related request and regulation.
2.4.3 repetitive test
Same eight taste dragon of batch is taken to bore 6 parts of particulate samples (lot number 20170501), respectively about 3g, accurately weighed, presses " 2.2.2 "
Method preparation under, measures under " 2.1 " item chromatographic condition, with the retention time and chromatographic peak area of No. 6 peaks (cucoline)
For reference, the relative retention time RSD that the relative retention time at remaining 11 shared peak in addition to cucoline is calculated exists
Between 0.18%~1.2%, relative peak area RSD is between 0.33%~1.8%, the results showed that and this method repeatability is good,
Meet the requirement of finger-print.
2.4.4 eight taste dragons bore the foundation of particle HPLC finger-print
Eight taste dragons are taken to bore particulate samples 10 batches, by legal system available test sample solution below " 2.2.2 " item, under " 2.1 " item
Analysis is measured under chromatographic condition, sees attached drawing 3.It establishes eight taste dragons and bores the common pattern map of particle HPLC finger-print (averagely
Value), it is analyzed using " similarity evaluation ", sees attached drawing 4.Symbiosis is at 12 on control map
A shared peak, wherein No. 6 chromatographic peak (cucoline) separating degrees are preferable in control map, and chromatography response is higher, and retention time occupies
In, therefore select No. 6 peaks (cucoline) for referring to peak.Eight taste dragons bore the evaluation of particle fingerprint similarity and use " Chinese medicine chromatography
Fingerprint similarity evaluation system (2012.130723 editions) " bores the testing number of the finger-print of particle to 10 batch, eight taste dragon
According to carry out processing analyze its similarity calculation the results are shown in Table 1.As shown in Table 1, the similarity of each eight taste dragon of batch brill particle is equal
0.989 or more, similitude is good.Show that contained index consistence of composition is preferable in each batch of sample.
1 10 batches of eight taste dragons of table bore the similarity of particle and reference fingerprint
2.4.5 the main common characteristic peaks attribution analysis of HPLC finger-print
It is accurate respectively to draw gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin and psoralen
Reference substance solution, by chromatographic condition analysis measurement under " 2.1 " item, recorded simultaneously using diode array detector chromatogram and
The ultra-violet absorption spectrum at each peak, and compared with each main shared peak in finger-print common pattern, as the result is shown
The retention time and its spectrum of 3,5,6,7,9,10, No. 12 characteristic peaks in finger-print common pattern respectively with gallic acid,
Protocatechuic acid, cucoline, rhodioside, catechin, the retention time and UV absorption light of aurantiamarin and psoralen reference substance
Spectrum is consistent, therefore No. 3 peaks for confirming that eight taste dragons bore particle map are gallic acid, and No. 5 peaks are protocatechuic acid, and No. 6 peaks are sinomenium acutum
Alkali, No. 7 peaks are rhodioside, and No. 9 peak catechins, No. 10 peaks are aurantiamarin, and No. 12 peaks are psoralen.
Claims (5)
1. a kind of can be used for detection method while eight taste dragons bore the quantification and qualification of particle main component, it is characterised in that
The method uses 70% methanol for solvent supersonic assisted extraction, is only referred to the HPLC that a chromatographic condition can measure said preparation simultaneously
The content of line map and 7 kinds of main components.
It is by toddalia, wammel, Radix zanthoxyli, caulis sinomenii, Caulis Spatholobi, nine 2. eight tastes dragon as described in claim 1 bores particle
The granule that Long Teng, alangium and eight taste medicine of radix fici simplicissimae are prepared.
3. the chromatographic condition of detection method as described in claim 1, measurement includes:
Using octadecylsilane chemically bonded silica as stationary phase, -0.1% phosphate aqueous solution of acetonitrile is mobile phase, gradient elution；Column temperature
It is 25~30 DEG C；Flow velocity is 1ml/min；Detector is UV detector, and Detection wavelength is respectively 270nm and 245nm.
4. detection method as described in claim 1, the chromatographic condition of measurement are as follows:
With Phenomenex Gemini C18(250mm × 4.6mm i.d., 5 μm) chromatographic column is stationary phase, acetonitrile (A) -0.1%
Phosphoric acid water (B) is mobile phase, and the percent by volume condition of gradient elution is: 0~10min, 6%A；10~20min, 12%A；
20~30min, 19%A；30~40min, 27%A；40~50min, 35%A；50~60min, 45%A；Column temperature is 25~30
℃；Flow velocity is 1ml/min；Detector is UV detector, and Detection wavelength is 0~50min:270nm, 50~60min:245nm；
10 μ L of sample volume；Chromatographic column column effect is not less than 30000 in terms of cucoline.
5. detection method as described in claim 1, the preparation of sample solution and measuring method include:
(1) preparation of reference substance solution: precision weighs gallic acid, protocatechuic acid, cucoline, rhodioside, catechin, orange peel
Glycosides and psoralen reference substance are appropriate, and methanol is added to dissolve, and debita spissitudo reference substance stock solution is respectively prepared.Separately take above-mentioned solution suitable
Amount, is placed in same measuring bottle, adds methanol dilution, is made containing 0.05~0.10mg/mL of gallic acid, containing protocatechuic acid and psoralea corylifolia
Plain concentration is 0.005~0.01mg/mL, is 0.1~0.3mg/mL containing cucoline, rhodioside, catechin concentration, contains aurantiamarin
Concentration is the mixed reference substance solution of 0.01~0.05mg/mL, is placed in freezer storage, spare；
(2) preparation of test solution: taking eight taste dragons to bore particle 3g, accurately weighed, sets in stuffed conical flask, 50mL is added in precision
70% methanol, weighed weight are ultrasonically treated (power 200W, frequency 40kHz) 20min, let cool, then weighed weight, with 70% first
Alcohol supplies the weight of less loss, shakes up, filtration, and subsequent filtrate centrifugation takes supernatant to get test solution；
(3) measuring method
It is accurate respectively to draw above-mentioned gallic acid containing a certain concentration, protocatechuic acid, cucoline, rhodioside, catechin, aurantiamarin
And each 10 μ l of mixed reference substance solution and test solution of psoralen, it is injected by chromatographic condition described in claim 3,4
High performance liquid chromatograph, measures corresponding peak area, calculate to get.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811226190.3A CN109374764B (en) | 2018-10-19 | 2018-10-19 | HPLC fingerprint spectrum and main component content determination method for eight-ingredient Longzui granules |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811226190.3A CN109374764B (en) | 2018-10-19 | 2018-10-19 | HPLC fingerprint spectrum and main component content determination method for eight-ingredient Longzui granules |
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| Publication Number | Publication Date |
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| CN109374764A true CN109374764A (en) | 2019-02-22 |
| CN109374764B CN109374764B (en) | 2021-05-14 |
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| CN114236034A (en) * | 2021-12-15 | 2022-03-25 | 贵州恒霸药业有限责任公司 | Detection method of toddalia asiatica medicinal material |
| CN114236034B (en) * | 2021-12-15 | 2024-03-29 | 贵州恒霸药业有限责任公司 | Method for detecting asiatic toddalia root blood medicinal material |
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