CN108562681B - The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent - Google Patents
The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent Download PDFInfo
- Publication number
- CN108562681B CN108562681B CN201810684983.3A CN201810684983A CN108562681B CN 108562681 B CN108562681 B CN 108562681B CN 201810684983 A CN201810684983 A CN 201810684983A CN 108562681 B CN108562681 B CN 108562681B
- Authority
- CN
- China
- Prior art keywords
- acid
- qiju dihuang
- reference substance
- dihuang koufuye
- koufuye
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent, the foundation of QIJU DIHUANG KOUFUYE multicomponent quantitative approach the following steps are included: step 1, QIJU DIHUANG KOUFUYE test solution preparation；The preparation of step 2, mixed reference substance solution: step 3, accurate absorption test solution and mixed reference substance solution inject liquid chromatograph respectively, record chromatogram；Step 4, the mixing reference substance obtained in step 3 and QIJU DIHUANG KOUFUYE form and aspect chromatogram are calculated into the content of each ingredient in sample according to retention time, peak area.QIJU DIHUANG KOUFUYE multicomponent quantitative analysis method provided by the present invention objectively can characterize the quality of QIJU DIHUANG KOUFUYE comprehensively.The quantitative detection method of multicomponent provided by the invention has method simplicity, stablizes, precision height, high repeatability and other advantages.
Description
Technical field
The present invention relates to a kind of detection methods of Chinese medicine, and in particular to the quantitative detection side of QIJU DIHUANG KOUFUYE multicomponent
Method.
Background technique
The corresponding treatment of Qiju Dihuang Wan is fire excess from yin deficiency, and the additional Qiju Dihuang Wan that clears liver and improve vision relatively nourishes the liver to improve visual acuity, and is suitable for liver
Kidney-yin deficiency, two blurred visions flower, blurred vision card.It is fire excess from yin deficiency to refer to that imbalance of yin and yang, the deficiency of Yin are then positive high and heat (at) fire, out
It swims outside yin asthenia generating intrinsic heat phenomenon, still there is flushing, the card such as hot eyes, dry throat, laryngalgia, bleeding, vexed, tongue fur is few, red tongue body is thin, thready rapid pulse.
It is fire excess from yin deficiency to refer to internal organs yin point virtual loss, it loses in performance raw in nourishing, abnormal heat.The heart, liver, lung, spleen, kidney may occur in which such feelings
Condition.Clinic is relatively conventional with kidney-yin deficiency, and symptom includes hectic fever night sweat, vexed, insomnia or man's premature ejaculation, spermatorrhea, woman through less or
Menostasis or vaporization heat of bone, soreness and weakness of waist and knees, tinnitus etc..Liver opening at eye, eyes can see with the hepatic blood injecting above.Fructus lycii tonifying kidney and benefiting sperm,
It nourishes the liver to improve visual acuity；Chrysanthemum is apt to head clearing, the heat of surname dissipate Liver Channel.The combination of eight kinds of compatibility of drugs is common to play enriching yin, nourishing the liver, improving eyesight
Act on nourishing the kidney and the liver.For liver-kidney Yin deficiency, dizziness and tinnitus, photophobia photophobia, tearing against wind, blurring of vision.
The content of single or several effective components is only measured about the Quality evaluation of QIJU DIHUANG KOUFUYE at present,
Cannot objective appraisal QIJU DIHUANG KOUFUYE comprehensively quality.The present invention establishes Lycium-rehmannia mouth using high performance liquid chromatography
The quantitative detecting method of 13 ingredients in liquid is taken, is medicinal material identification, quality evaluation and the quality mark of QIJU DIHUANG KOUFUYE
Quasi- formulation has great importance.
Summary of the invention
Goal of the invention: more present invention aims to solve the deficiencies of the prior art, and provides a kind of a kind of QIJU DIHUANG KOUFUYE
Component quantifying detection method, which can objective, comprehensively and accurately evaluate the quality of QIJU DIHUANG KOUFUYE, to control
The quality of QIJU DIHUANG KOUFUYE processed and guarantee that clinical efficacy is of great significance.
Technical solution: to achieve the goals above, the technical scheme adopted by the invention is as follows:
A kind of detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative, which comprises the following steps:
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution:
QIJU DIHUANG KOUFUYE is taken, precision is measured, set in evaporating dish, and water-bath volatilizes, and methanol is added and is ultrasonically treated (power
150W, frequency 20kHz) 30 minutes, let cool, filter to obtain clear liquid, constant volume, cross 0.45 μm of miillpore filter to get.
The preparation of step 2, acids mixed reference substance solution:
Chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B, appropriate Paeonol are taken, essence
It is close weighed, add methanol that mixed reference substance solution is made, shakes up.
Step 3, the preparation of saponins mixed reference substance solution take 5 hydroxymethyl furfural (5-HMF), morroniside, loganin, Chinese herbaceous peony
Medicine glycosides, acteoside, appropriate luteolin, it is accurately weighed, add methanol that every 1mL 80ug containing 5-HMF, morroniside is made
The mixed reference substance solution of 120ug, loganin 10ug, Paeoniflorin 300ug, acteoside 10ug, luteolin 30ug, shake
It is even；
Step 4, respectively accurate absorption test solution and control solution, inject high performance liquid chromatograph, record chromatography
Figure；
Step 5, by the mixing reference substance obtained in step 4 and QIJU DIHUANG KOUFUYE form and aspect chromatogram, when according to retaining
Between, peak area, calculate sample in each ingredient content.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, step 1 Qi chrysanthemum
Rehmannia-root oral liquid sample solution preparation method are as follows: take 3 batches takes QIJU DIHUANG KOUFUYE 10mL, and precision measures, and sets evaporation
In ware, water-bath is volatilized, and 5ml methanol ultrasonic treatment (power 150W, frequency 20kHz) is added 30 minutes, is let cool, is filtered to obtain clear liquid,
It is settled to 10mL, crosses 0.45 μm of miillpore filter to get test solution.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, step 2 acids
The preparation of mixed reference substance solution: take chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B,
Appropriate Paeonol, it is accurately weighed, add methanol be made every 1mL 90ug containing chlorogenic acid, Cryptochlorogenic acid 60ug, 3,4-Dicaffeoylquinic acid 10ug,
The mixed reference substance solution of 3,5-Dicaffeoylquinic acid 50ug, 4,5-Dicaffeoylquinic acid 50ug, tanshin polyphenolic acid B 40ug, Paeonol 50ug, shake up,
?.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, step 3 saponin(e
The preparation of class mixed reference substance solution takes 5-HMF, morroniside, loganin, Paeoniflorin, acteoside, appropriate luteolin, accurate
It is weighed, add methanol that every 1mL 80ug containing 5-HMF, morroniside 120ug, loganin 10ug, Paeoniflorin 300ug, verbascose is made
The mixed reference substance solution of glycosides 10ug, luteolin 30ug, shake up.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, in step 4, liquid
Phase chromatographic condition are as follows: liquid phase chromatogram condition are as follows: chromatographic column: YMC-Pack ODS-A (250 × 4.6mm, 5um, 12nm)；Flowing
It is mutually 0.1% formic acid-acetonitrile solution, gradient elution (such as table 1), diode array detector (DAD) 210-800nm all-wave length is swept
It retouches, Detection wavelength: 254nm and 325nm, 35 DEG C of column temperature, flow velocity 1mL/min, sampling volume: 10uL.
1 gradient elution program of table
| Program | Time (min) | Acetonitrile (%) | 0.1% aqueous formic acid (%) |
| 1 | 0 | 5 | 95 |
| 2 | 10 | 5 | 95 |
| 3 | 20 | 10 | 90 |
| 4 | 90 | 30 | 70 |
| 7 | 105 | 55 | 45 |
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, which is characterized in that
The content of 13 ingredients such as phenolic acid class and saponins is detected simultaneously.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, it is step 5, accurate
Draw phenolic acid class mixed reference substance solution and each 1,2,4,8,10,15,20 μ L sample introduction of saponins hybrid standard product solution, measurement
Peak area, with reference substance sample volume x (μ g) for abscissa, peak area y is ordinate, draws standard curve, calculates regression equation:
Standard curve
The optimization of multicomponent quantitative detection condition:
1, in terms of the preparation optimization of sample solution
The present invention is by using different solvents (methanol, water, 70% second after the processing such as being evaporated, diluting, extract to sample
Alcohol solution, 85% ethanol water, 95% ethyl alcohol, dehydrated alcohol) it redissolves, carry out experiment comparison, as a result, it has been found that multiple after being evaporated
Molten resulting spectrogram differential response value is higher, and the limit is steady, therefore using the method redissolved after being evaporated；To in the investigation for redissolving solvent
It was found that methanolic extract chromatogram information content is most, component content highest；So water-bath is selected to volatilize, methanol is added to be ultrasonically treated.
2, aspect is optimized in chromatographic condition
The present invention investigates Detection wavelength using diode array detector, extract 254nm, 280nm, 284nm,
Chromatogram at 300nm, 325nm, as a result, it has been found that when Detection wavelength is 254nm, information that saponin component chromatogram is included
Amount is most comprehensive and baseline is steady, therefore selecting 254nm is saponin component Detection wavelength；It was found that when Detection wavelength is 325nm, phenolic acid class
The information content that ingredient chromatogram is included is most comprehensive and baseline is steady, therefore selecting 325nm is liposoluble ingredient Detection wavelength；
The present invention sieves flow velocity (1ml/min, 0.8ml/min, 0.7ml/min, 0.6ml/min, 0.5ml/min)
Choosing, because more there is isomer and the very much like ingredient of other polarity, Gu Gaoliu in the ingredient in QIJU DIHUANG KOUFUYE
It can not be separated under speed, therefore separating effect is preferable at low flow rates, finally under the conditions of flow velocity is more constant gradients of 1ml/min
The similar substance of polarity is separated.
The present invention compares methanol-water, acetonitrile-water, -0.1% formic acid of acetonitrile, acetonitrile and 0.05% phosphoric acid water, acetonitrile -
Elution effect of the 0.1% phosphoric acid water 5 difference elution system under different gradients.As a result, it has been found that with acetonitrile and 0.1% formic acid water,
When for mobile phase, each ingredient can reach good separating effect in QIJU DIHUANG KOUFUYE, therefore final selected with acetonitrile and 0.1%
Formic acid water is mobile phase.
The utility model has the advantages that
1, the structural property feature of the active constituent of the invention according to contained in QIJU DIHUANG KOUFUYE, passes through many experiments
Filter out optimal flowing phase composition, gradient elution program, flow velocity, Detection wavelength, chromatographic column, the analysis conditions such as column temperature, through more
Secondary experimental verification shows QIJU DIHUANG KOUFUYE multicomponent quantitative detecting method provided by the invention, can be comprehensive, objective, quasi-
The quality of true detection and evaluation QIJU DIHUANG KOUFUYE, to guarantee that clinical QIJU DIHUANG KOUFUYE curative effect is of great significance.
2, the QIJU DIHUANG KOUFUYE multicomponent established with method provided by the present invention is quantitative, can effectively characterize Qi
The quality of Ju Dihuang oral solution objective can embody each tandem and correlation for constituting fingerprint characteristic peak, focus on whole
Facial feature not only can avoid determining the one-sidedness of QIJU DIHUANG KOUFUYE quality because measuring individual chemical ingredient, but also can reduce
A possibility that artificially being handled for requisite quality.
3, the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent provided by the invention has method simplicity, stability
Good, precision height, high repeatability and other advantages.
Detailed description of the invention
Fig. 1 is the chromatogram of saponins reference substance solution.
Fig. 2 is the chromatogram of phenolic acid class reference substance solution.
Fig. 3 is chromatogram of the QIJU DIHUANG KOUFUYE test sample at 254nm.
Fig. 4 is the chromatogram of QIJU DIHUANG KOUFUYE test sample at 325 nm.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, actual conditions are not specified in embodiment
Person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer, being can be with
Conventional products that are commercially available.
The instrument that embodiment is used and reagent are as follows:
Experiment equipment
1.1 instrument
Japanese Shimadzu Corporation's all band scans (200-800nm) high liquid chromatography system, including full-automatic on-line degassing system
System, full-automatic sampling system Prominence SIL-20A, diode array detector SPD-M20A and automatic temperature-controlled column oven
CTO-20A, KQ3200DB type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), BP121S electronic analytical balance
(SARTORIUS)。
1.2 drug and reagent
QIJU DIHUANG KOUFUYE sample be all from Jiangsu Hai Sheng pharmaceutcal corporation, Ltd (sample lot number is respectively as follows: 140401,
160301,160503)；5-HMF reference substance (lot number: 111626-201509), morroniside reference substance (lot number: 111998-
201501, purity: 96.8%), chlorogenic acid reference substance (lot number: 110753-201415, purity: 96.2%), Cryptochlorogenic acid control
, Chinese herbaceous peony product (lot number: 150728 purity: >=98%), loganin reference substance (lot number: 111640-201005, purity: 99.2%)
Glycosides reference substance (lot number: 110736-201539), acteoside reference substance (lot number: 61276-17-3, purity: >=98%) southern
, different green Jing Senbeijia Biotechnology Co., Ltd, luteolin reference substance (lot number: 111720-200905, purity: 96.5%)
Ortho acid B reference substance (lot number: 150726), 3,5-Dicaffeoylquinic acid reference substance (lot number: 151028), 4,5-Dicaffeoylquinic acid reference substance (lot number:
150624), tanshin polyphenolic acid B reference substance (lot number: 111562-201615), Paeonol reference substance (lot number: 110708-200506, it is pure
Degree: >=98%) it is purchased from Chinese pharmaceutical biological product inspection institute；Methanol (analysis is pure)；Petroleum ether (analysis is pure)；Ether (analysis
It is pure)；Phosphoric acid (analysis is pure)；Acetonitrile (chromatographically pure)；The used reagent of experiment is purchased from Nanjing Chemistry Reagent Co., Ltd., Wa Ha
Breathe out pure water.
A kind of detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative of embodiment 1, comprising the following steps:
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution:
3 crowdes of QIJU DIHUANG KOUFUYE 10mL of the above table 1 are taken, precision is measured, set in evaporating dish, and water-bath volatilizes, and 5ml is added
Methanol is ultrasonically treated (power 150W, frequency 20kHz) 30 minutes, lets cool, filters to obtain clear liquid, is settled to 10mL, and 0.45 μm excessively micro-
Hole filter membrane is to get test solution.
The preparation of step 2, acids mixed reference substance solution:
Chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B, appropriate Paeonol are taken, essence
It is close weighed, add methanol that every 1mL 90ug containing chlorogenic acid, Cryptochlorogenic acid 60ug, 3,4-Dicaffeoylquinic acid 10ug, 3,5-Dicaffeoylquinic acid is made
The mixed reference substance solution of 50ug, 4,5-Dicaffeoylquinic acid 50ug, tanshin polyphenolic acid B 40ug, Paeonol 50ug, shake up.
Step 3, the preparation of saponins mixed reference substance solution take 5-HMF, morroniside, loganin, Paeoniflorin, verbascose
Glycosides, appropriate luteolin, it is accurately weighed, add methanol be made every 1mL 80ug containing 5-HMF, morroniside 120ug, loganin 10ug,
The mixed reference substance solution of Paeoniflorin 300ug, acteoside 10ug, luteolin 30ug, shake up.
Step 4, respectively 3 batches of QIJU DIHUANG KOUFUYE test solution and control solution of accurate absorption, inject efficient liquid
Chromatography records chromatogram, mixes reference substance chromatography such as Fig. 1 and Fig. 2, the chromatogram of test solution such as Fig. 3 and Fig. 4；(figure
Middle peak 1 is 5-HMF；Peak 2 is morroniside；Peak 3 is chlorogenic acid；Peak 4 is Cryptochlorogenic acid；Peak 5 is loganin；Peak 6 is Paeoniflorin；Peak
7 be acteoside；Peak 8 is luteolin；Peak 9 is 3,4-Dicaffeoylquinic acid；Peak 10 is 3,5-Dicaffeoylquinic acid；Peak 11 is 4,5-Dicaffeoylquinic acid；Peak
12 be tanshin polyphenolic acid B；Peak 13 is Paeonol) chromatographic column: YMC-Pack ODS-A (250 × 4.6mm, 5um, 12nm)；Mobile phase is
0.1% formic acid-acetonitrile solution, gradient elution such as table 1, diode array detector (DAD) 210-800nm full wavelength scanner, inspection
Survey wavelength: 254nm and 325nm, 35 DEG C of column temperature, flow velocity 1mL/min, sampling volume: 10uL.
1 gradient elution program of table
1, linear relationship is investigated
Take phenolic acid class mixed reference substance solution and each 1,2,4,8,10,15,20 μ L of saponins hybrid standard product solution into
Sample measures peak area, and with reference substance sample volume x (μ g) for abscissa, peak area y is ordinate, draws standard curve, calculates back
Return equation.It the results are shown in Table 2.
2 standard curve of table
2 methodological study of embodiment
1, precision research
Sample lot number by the preparation of 1 method of embodiment is 140401 test solutions, according to the detection method point of embodiment 1
Analysis, parallel sample introduction 6 times, sample volume is 10 μ L, with 5-HMF, morroniside, chlorogenic acid, Cryptochlorogenic acid, loganin, Paeoniflorin, Mao Rui
Flower glucosides, luteolin, RSD (%) value of 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B, Paeonol peak area
Respectively 1.32,2.58,3.45,2.01,1.94,1.53,1.49,2.05,2.53,2.41,0.98,2.40,2.58, as a result
Show that the parallel sample introduction accuracy of the equipment is good.
2, stability study
Sample lot number by the preparation of 1 method of embodiment is 140401 test solutions, according to the detection method point of embodiment 1
Analysis, using 0,2,6,12,18,24 hour different time, sample introduction is analyzed, and sample volume is 10 μ L, with 5-HMF, morroniside, green original
Acid, Cryptochlorogenic acid, loganin, Paeoniflorin, acteoside, luteolin, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid,
Tanshin polyphenolic acid B, Paeonol peak area RSD (%) value be respectively 2.47,1.25,2.53,3.47,3.15,2.34,1.13,
1.14,1.83,2.64,2.89,2.58,2.79, the results showed that chromatographic peak area variation of the test solution within for 24 hours is not
Greatly, stability is very good.
3, repetitive test
It is 140401 test solutions that parallel precision, which weighs lot number, is analyzed according to the detection method of embodiment 1, sample volume is
10 μ L, with 5-HMF, morroniside, chlorogenic acid, Cryptochlorogenic acid, loganin, Paeoniflorin, acteoside, luteolin, different green original
Sour B, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B, Paeonol peak area RSD (%) value be respectively 1.72,1.32,2.35,
2.05,1.52,1.63,1.42,2.43,2.38,1.05,1.86,2.56,2.35, the results showed that sample chromatogram peak reproducibility
Good, the repeatability of this method is good.
Above the experimental results showed that, QIJU DIHUANG KOUFUYE multicomponent quantitative detecting method provided by the invention, stability
Good, precision is high, reproducible, can objectively evaluate the quality of QIJU DIHUANG KOUFUYE comprehensively, to guarantee that clinical efficacy has weight
The meaning wanted.
Above embodiments are only exemplary embodiment of the present invention, are not used in the limitation present invention, protection scope of the present invention
It is defined by the claims.Those skilled in the art can within the spirit and scope of the present invention make respectively the present invention
Kind modification or equivalent replacement, this modification or equivalent replacement also should be regarded as being within the scope of the present invention.
Claims (1)
1. a kind of detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative, which comprises the following steps:
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution:
3 different batches QIJU DIHUANG KOUFUYE 10mL are taken, precision is measured, set in evaporating dish, and water-bath volatilizes, and 5ml methanol is added
Ultrasonic treatment is let cool for 30 minutes, ultrasonic power 150W, frequency 20kHz, filters to obtain clear liquid, is settled to 10mL, crosses 0.45 μm of micropore
Filter membrane to get；
The preparation of step 2, acids mixed reference substance solution:
Chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B and appropriate Paeonol are taken, it is accurate
It is weighed, add methanol be made every 1mL containing 90 μ g of chlorogenic acid, 60 μ g of Cryptochlorogenic acid, 10 μ g of 3,4-Dicaffeoylquinic acid, 50 μ g of 3,5-Dicaffeoylquinic acid,
The mixed reference substance solution of 50 μ g of 4,5-Dicaffeoylquinic acid, 40 μ g of tanshin polyphenolic acid B, 50 μ g of Paeonol, shake up；
Step 3, the preparation of saponins mixed reference substance solution:
5 hydroxymethyl furfural, morroniside, loganin, Paeoniflorin, acteoside, appropriate luteolin are taken, it is accurately weighed, add first
Every 1mL is made containing 80 μ g of 5 hydroxymethyl furfural, 120 μ g of morroniside, 10 μ g of loganin, 300 μ g of Paeoniflorin, acteoside 10 in alcohol
The mixed reference substance solution of μ g, 30 μ g of luteolin, shake up；
Step 4, respectively accurate absorption test solution and control solution, inject high performance liquid chromatograph, record chromatogram；
Step 5, by the liquid chromatogram of the mixing reference substance and QIJU DIHUANG KOUFUYE test sample that are obtained in step 4, according to guarantor
Time, peak area are stayed, the content of each ingredient in QIJU DIHUANG KOUFUYE test sample is calculated；
In step 4, the chromatographic condition of high performance liquid chromatograph are as follows: chromatographic column: YMC-Pack ODS-A, 250 × 4.6mm, 5 μm,
12nm；Mobile phase is 0.1% formic acid and acetonitrile solution, and gradient elution, diode array detector, 210-800nm all-wave length sweeps
It retouches, Detection wavelength: 254nm and 325nm, 35 DEG C of column temperature, flow velocity 1mL/min, sampling volume: 10 μ L；
Gradient elution program is as follows:
0~10 minute: acetonitrile volume 5~5%, 0.1% aqueous formic acid volume 95~95%；
10~20 minutes: acetonitrile volume 5~10%, 0.1% aqueous formic acid volume 95~90%；
20~90 minutes: acetonitrile volume 10~30%, 0.1% aqueous formic acid volume 90~70%；
90~105 minutes: acetonitrile volume 30~55%, 0.1% aqueous formic acid volume 70~45%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810684983.3A CN108562681B (en) | 2018-06-28 | 2018-06-28 | The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810684983.3A CN108562681B (en) | 2018-06-28 | 2018-06-28 | The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108562681A CN108562681A (en) | 2018-09-21 |
| CN108562681B true CN108562681B (en) | 2019-04-05 |
Family
ID=63554881
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810684983.3A Active CN108562681B (en) | 2018-06-28 | 2018-06-28 | The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108562681B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113125569B (en) * | 2019-12-30 | 2022-07-22 | 湖南易能生物医药有限公司 | Fingerprint spectrum determination method and quality control method based on Guyinshi material |
| CN111830160B (en) * | 2020-07-21 | 2022-06-14 | 上海和黄药业有限公司 | Method for detecting fingerprint spectrum of Qijudihuang pills and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1600364A (en) * | 2004-10-12 | 2005-03-30 | 江西江中药业股份有限公司 | Method for controlling quality of oral liquid of medlar-chrysanthemum-rehmannia root |
| CN102768242A (en) * | 2011-04-30 | 2012-11-07 | 顾建光 | Method for determining content of oral liquid of Chinese wolfberry and rehmanniae |
| CN106177345A (en) * | 2016-09-06 | 2016-12-07 | 上海辰松新材料科技有限公司 | QIJU DIHUANG WAN and preparation method thereof |
| CN107648424A (en) * | 2017-11-07 | 2018-02-02 | 贵州拜特制药有限公司 | A kind of preparation method of Lycium-rehmannia preparation |
-
2018
- 2018-06-28 CN CN201810684983.3A patent/CN108562681B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1600364A (en) * | 2004-10-12 | 2005-03-30 | 江西江中药业股份有限公司 | Method for controlling quality of oral liquid of medlar-chrysanthemum-rehmannia root |
| CN102768242A (en) * | 2011-04-30 | 2012-11-07 | 顾建光 | Method for determining content of oral liquid of Chinese wolfberry and rehmanniae |
| CN106177345A (en) * | 2016-09-06 | 2016-12-07 | 上海辰松新材料科技有限公司 | QIJU DIHUANG WAN and preparation method thereof |
| CN107648424A (en) * | 2017-11-07 | 2018-02-02 | 贵州拜特制药有限公司 | A kind of preparation method of Lycium-rehmannia preparation |
Non-Patent Citations (4)
| Title |
|---|
| HPLC法同时测定不同产地不同品种菊花中8种成分的含量;屠万倩等;《中药材》;20180131;第41卷(第1期);第147页左栏第1段，第2.2节 |
| RP-HPLC法测定六味地黄丸中丹皮酚、芍药苷和乌苏酸;李向阳等;《中成药》;20120228;第34卷(第2期);第278页第2段，第2.1.2节 |
| 一测多评法同时测定杞菊地黄口服液中4种有效成分的含量;王晓燕等;《药物分析杂志》;20170228;第37卷(第2期);第2.1.1节，第2.1.2节，第2.3.1节，第4节，第291页左栏第1段 |
| 鲜、生、熟地黄药材中3种活性成分含量的比较;王宏洁等;《中国中药杂志》;20080831;第33卷(第15期);第1923页左栏第1段，第2.3节 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108562681A (en) | 2018-09-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105842373B (en) | A method of establishing the finger-print of the pharmaceutical preparation of honeysuckle | |
| CN109709251A (en) | The detection method of linggui zhugan decoction finger-print | |
| CN110031570A (en) | The fingerprint atlas detection method of Kangganmao Granule | |
| CN108226313A (en) | In glutinous rehmannia while methods of glycosides measure and fingerprint map construction method | |
| CN106198782A (en) | A kind of can realize the content analysis of 18 components in Folium Ilicis and the method for quality control of similarity evaluation simultaneously | |
| CN108562681B (en) | The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent | |
| CN109633001A (en) | The fingerprint atlas detection method of the wine fruit of glossy privet | |
| CN110702813A (en) | Miao medicine caulis et folium periplocae HPLC fingerprint spectrum research and multi-component content determination method | |
| CN108254470B (en) | Method for simultaneously measuring saccharide components in rehmannia and establishing fingerprint spectrum of saccharide components | |
| CN110441441A (en) | A kind of the UPLC characteristic spectrum construction method and discrimination method of rhizoma cibotii and boiling hot rhizoma cibotii medicinal material | |
| CN109187799A (en) | A kind of fingerprint and its standard finger-print of perilla seed standard decoction | |
| CN105891403A (en) | Anemone flaccida medicinal material HPLC-UV characteristic spectrum construction method | |
| CN108802233A (en) | The detection method of a variety of effective components in a kind of Chinese cassia tree | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN104807932A (en) | Determining method of fingerprint spectrum of flavonoid ingredient of fenugreek seed extract | |
| CN108020617A (en) | A kind of method of Nitidine Chloride content in measure shiny pricklyash toothpaste | |
| CN104597168B (en) | Ramulus Mori refines decoction pieces content assaying method | |
| CN107064325A (en) | A kind of method of quality control of Qige granules | |
| CN105353065A (en) | Establishing method of HPLC (high-performance liquid chromatography) fingerprint spectrum of lychee seeds, standard fingerprint spectrum obtained with method and application of standard fingerprint spectrum | |
| CN110118841A (en) | A kind of construction method of the HPLC characteristic spectrum of oral liquid for clearing liver and gallbladder | |
| CN109655542A (en) | A kind of construction method of Oroxylum indicum granule UPLC characteristic spectrum and its application | |
| CN110361459A (en) | The method for building up of loguat leaf standard decoction finger-print | |
| CN109521119A (en) | A kind of measuring method of compound south isatis root granules finger-print | |
| CN110763642B (en) | Detection method of traditional Chinese medicine preparation | |
| CN107748217B (en) | A kind of method for building up of Pisces particle GC quantitative finger print atlas and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |