CN108562681A - The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent - Google Patents
The quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent Download PDFInfo
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G—PHYSICS
- 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
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Abstract
The invention discloses the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent, the foundation of QIJU DIHUANG KOUFUYE multicomponent quantitative approach includes the following steps：The preparation of step 1, QIJU DIHUANG KOUFUYE test solution；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 easy method, stable, 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 technology
Qiju Dihuang Wan corresponds to the fire excess from yin deficiency additional Qiju Dihuang Wan that clears liver and improve vision for the treatment of and relatively nourishes the liver to improve visual acuity, and is suitable for liver
The card of kidney-yin deficiency, two blurred visions flower, blurred vision.Fire excess from yin deficiency refers to that imbalance of yin and yang, the deficiency of Yin are then positive high and heat (at) is fiery, is gone out
It swims outside yin asthenia generating intrinsic heat phenomenon, still there is flushing, the cards such as hot eyes, dry throat, laryngalgia, bleeding, vexed, tongue is few, red tongue body is thin, thready rapid pulse.
Fire excess from yin deficiency refers to internal organs the moon point virtual loss, is lost 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, seminal emission, woman through less or
Amenorrhea 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.Fruit of Chinese wolfberry 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 active ingredients 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 discriminating, quality evaluation and the quality mark of QIJU DIHUANG KOUFUYE
Accurate formulation has great importance.
Invention content
Goal of the invention：It is 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 ensure that clinical efficacy is of great significance.
Technical solution：To achieve the goals above, the technical solution that the present invention takes is：
A kind of detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative, which is characterized in that include the following steps：
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution：
QIJU DIHUANG KOUFUYE, precision is taken to measure, set in evaporating dish, 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, you can.
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, you can；
Step 4, respectively accurate absorption test solution and reference substance 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
Yellow oral solution test solution preparation method is：Take 3 batches takes QIJU DIHUANG KOUFUYE 10mL, precision to measure, and sets evaporating dish
In, water-bath volatilizes, and 5ml methanol is added and is ultrasonically treated (power 150W, frequency 20kHz) 30 minutes, lets cool, filters to obtain clear liquid, fixed
Hold 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 are mixed
Close the preparation of reference substance solution：Take chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, tanshin polyphenolic acid B, pellet
Skin phenol is appropriate, accurately weighed, adds methanol that every 1mL 90ug containing chlorogenic acid, Cryptochlorogenic acid 60ug, 3,4-Dicaffeoylquinic acid 10ug, different are made
The mixed reference substance solution of chlorogenic acid A 50ug, 4,5-Dicaffeoylquinic acid 50ug, tanshin polyphenolic acid B 40ug, Paeonol 50ug, shake up, i.e.,
It can.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, step 3 saponins
Mixed reference substance solution preparation takes 5-HMF, morroniside, loganin, Paeoniflorin, acteoside, appropriate luteolin, precision to claim
It is fixed, add methanol that every 1mL 80ug containing 5-HMF, morroniside 120ug, loganin 10ug, Paeoniflorin 300ug, acteoside is made
The mixed reference substance solution of 10ug, luteolin 30ug, shake up, you can.
Preferably, the quantitative detection method of above-described QIJU DIHUANG KOUFUYE multicomponent, in step 4, liquid
Phase chromatographic condition is：Liquid phase chromatogram condition is：Chromatographic column：YMC-Pack ODS-A (250 × 4.6mm, 5um, 12nm)；Mobile phase
For 0.1% formic acid-acetonitrile solution, gradient elution (such as table 1), diode array detector (DAD) 210-800nm all-wave lengths are swept
Face, Detection wavelength：254nm and 325nm, 35 DEG C, flow velocity 1mL/min of column temperature, 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
Phenolic acid class mixed reference substance solution and each 1,2,4,8,10,15,20 μ L sample introductions of saponins hybrid standard product solution are drawn, peak is measured
Area, with reference substance sample size x (μ g) for abscissa, peak area y is ordinate, draws standard curve, calculates regression equation:
Standard curve
Multicomponent quantifies the optimization of testing conditions：
1, in terms of the preparation optimization of sample solution
The present invention after the processing such as being evaporated, diluting, extract to sample by using different solvents (methanol, water, 70% second
Alcohol solution, 85% ethanol water, 95% ethyl alcohol, absolute ethyl alcohol) it redissolves, carry out experiment comparison, as a result, it has been found that multiple after being evaporated
The spectrogram differential response value of molten gained is higher, and the limit is steady, therefore using the method redissolved after being evaporated；In investigation to 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, extraction 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 it is saponin component Detection wavelength to select 254nm；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 it is liposoluble ingredient Detection wavelength to select 325nm；
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,
For mobile phase when, 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.
Advantageous effect：
1, the present invention passes through many experiments according to the structural property feature of active constituent contained in QIJU DIHUANG KOUFUYE
Filter out best 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, accurate
The quality of true detection and evaluation QIJU DIHUANG KOUFUYE, to ensure 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 judging the one-sidedness of QIJU DIHUANG KOUFUYE quality because measuring individual chemical ingredient, but also can reduce
The possibility artificially 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.
Description of the drawings
Fig. 1 is the chromatogram of phenolic acid class reference substance solution.
Fig. 2 is the chromatogram of saponins 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 implementation mode
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 balances
(SARTORIUS)。
1.2 drugs and reagent
QIJU DIHUANG KOUFUYE sample is all from Jiangsu Haisheng Pharmaceutical Co., Ltd., and (sample lot number is respectively：140401、
160301、160503)；5-HMF reference substance (lot numbers：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 compares
Product (lot number：150728 purity：>=98%), loganin reference substance (lot number：111640-201005, purity：99.2%), Chinese herbaceous peony
Glycosides reference substance (lot number：110736-201539), acteoside reference substance (lot number：61276-17-3, purity：>=98%) southern
The bio tech ltd Jing Senbeijia, luteolin reference substance (lot number：111720-200905, purity：96.5%), different green
Ortho acid B reference substance (lot numbers：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 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 1 detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative of embodiment, includes the following steps：
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution：
3 crowdes of QIJU DIHUANG KOUFUYE 10mL with upper table 1, precision are taken to measure, set in evaporating dish, 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 be made every 1mL 90ug containing chlorogenic acid, Cryptochlorogenic acid 60ug, 3,4-Dicaffeoylquinic acid 10ug, 3,5-Dicaffeoylquinic acid 50ug,
The mixed reference substance solution of 4,5-Dicaffeoylquinic acid 50ug, tanshin polyphenolic acid B 40ug, Paeonol 50ug, shake up, you can.
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, you can.
Step 4, respectively 3 batches of QIJU DIHUANG KOUFUYE test solutions of accurate absorption and reference substance solution, inject efficient liquid
Chromatography records chromatogram, mixes reference substance chromatography such as Fig. 1 and Fig. 2, chromatogram such as Fig. 3 and Fig. 4 of test solution；(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 all-wave length surface sweepings, inspection
Survey wavelength：254nm and 325nm, 35 DEG C, flow velocity 1mL/min of column temperature, sampling volume：10uL.
1 gradient elution program of table
1, linear relationship is investigated
Phenolic acid class mixed reference substance solution and each 1,2,4,8,10,15,20 μ L sample introductions of saponins hybrid standard product solution are taken,
Peak area is measured, with reference substance sample size x (μ g) for abscissa, peak area y is ordinate, draws standard curve, calculates and returns
Equation.It the results are shown in Table 2.
2 standard curve of table
2 methodological study of embodiment
1, precision research
It is 140401 test solutions by sample lot number prepared by 1 method of embodiment, according to the detection method point of embodiment 1
Analysis, parallel sample introduction 6 times, sample size is 10 μ L, with 5-HMF, morroniside, chlorogenic acid, Cryptochlorogenic acid, loganin, Paeoniflorin, Mao Rui
RSD (%) value of flower glucosides, luteolin, 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 table
The parallel sample introduction accuracy of the bright equipment is good.
2, stability study
It is 140401 test solutions by sample lot number prepared by 1 method of embodiment, according to the detection method point of embodiment 1
Analysis, using 0,2,6,12,18,24 hour different time, sample introduction is analyzed, sample size be 10 μ L, with 5-HMF, morroniside, chlorogenic acid,
Cryptochlorogenic acid, loganin, Paeoniflorin, acteoside, luteolin, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, red phenol
Sour 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 little, stablizes
Property 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 size 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 favorable reproducibility,
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, the quality of the comprehensive objective evaluation QIJU DIHUANG KOUFUYE of energy, to ensure that clinical efficacy has weight
The meaning wanted.
Above example is only exemplary embodiment of the present invention, is 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 (6)
1. a kind of detection method that QIJU DIHUANG KOUFUYE multicomponent is quantitative, which is characterized in that include the following steps：
The preparation of step 1, QIJU DIHUANG KOUFUYE test solution：
QIJU DIHUANG KOUFUYE, precision is taken to measure, set in evaporating dish, water-bath volatilizes, and methanol is added and is ultrasonically treated, lets cool, filters
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 and appropriate Paeonol are taken, it is accurate
It is weighed, add methanol that mixed reference substance solution is made, shakes up, you can；
It is prepared by step 3, saponins mixed reference substance solution：
5 hydroxymethyl furfural, morroniside, loganin, Paeoniflorin, acteoside and appropriate luteolin are taken, it is accurately weighed, add
Mixed reference substance solution is made in methanol, shakes up, you can；
Step 4, respectively accurate absorption test solution and reference substance 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.
2. the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent according to claim 1, which is characterized in that step 1
QIJU DIHUANG KOUFUYE test solution preparation method is：3 different batches QIJU DIHUANG KOUFUYE 10mL, precision are taken to measure,
It sets in evaporating dish, water-bath volatilizes, and 5ml methanol supersound process is added and lets cool, mistake within 30 minutes, ultrasonic power 150W, frequency 20kHz
Filter to obtain clear liquid, be settled to 10mL, cross 0.45 μm of miillpore filter to get.
3. the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent according to claim 1, which is characterized in that step 2
The preparation of phenolic acid class mixed reference substance solution：Take chlorogenic acid, Cryptochlorogenic acid, 3,4-Dicaffeoylquinic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, pellet
Phenolic acid B and appropriate Paeonol, it is accurately weighed, add methanol that every 1mL 90ug containing chlorogenic acid, Cryptochlorogenic acid 60ug, 3,4-Dicaffeoylquinic acid is made
The mixed reference substance solution of 10ug, 3,5-Dicaffeoylquinic acid 50ug, 4,5-Dicaffeoylquinic acid 50ug, tanshin polyphenolic acid B 40ug, Paeonol 50ug, shake
It is even, you can.
4. the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent according to claim 1, which is characterized in that step 3,
It is prepared by saponins mixed reference substance solution：Take 5 hydroxymethyl furfural, morroniside, loganin, Paeoniflorin, acteoside, wooden slippers
Careless glycosides is appropriate, accurately weighed, adds methanol that every 1mL 80ug containing 5 hydroxymethyl furfural, morroniside 120ug, loganin 10ug, Chinese herbaceous peony is made
The mixed reference substance solution of medicine glycosides 300ug, acteoside 10ug, luteolin 30ug, shake up, you can.
5. the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent according to claim 1, which is characterized in that step 4
In, the chromatographic condition of high performance liquid chromatograph is：Chromatographic column：YMC-Pack ODS-A, 250 × 4.6mm, 5um, 12nm；Flowing
It is mutually 0.1% formic acid and acetonitrile solution, gradient elution, diode array detector (DAD) 210-800nm all-wave length surface sweepings, inspection
Survey wavelength：254nm and 325nm, 35 DEG C, flow velocity 1mL/min of column temperature, sampling volume：10uL；
Gradient elution program such as following table：
6. the quantitative detection method of QIJU DIHUANG KOUFUYE multicomponent according to claim 1, which is characterized in that step 5,
Precision draws phenolic acid class mixed reference substance solution and each 1,2,4,8,10,15,20 μ L sample introductions of saponins hybrid standard product solution, surveys
Determine peak area, with reference substance sample size x (μ g) for abscissa, peak area y is ordinate, draws standard curve, calculates recurrence side
Journey:
Regression equation
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|---|---|---|---|---|
| CN111830160A (en) * | 2020-07-21 | 2020-10-27 | 上海和黄药业有限公司 | Method for detecting fingerprint spectrum of Qijudihuang pills and application thereof |
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| 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 |
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