CN108802229A - The screening technique of the processing procedure of wine glutinous rehmannia - Google Patents
The screening technique of the processing procedure of wine glutinous rehmannia Download PDFInfo
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- CN108802229A CN108802229A CN201810641930.3A CN201810641930A CN108802229A CN 108802229 A CN108802229 A CN 108802229A CN 201810641930 A CN201810641930 A CN 201810641930A CN 108802229 A CN108802229 A CN 108802229A
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- martynoside
- catalpol
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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
Abstract
The invention discloses the screening technique of the processing procedure of wine glutinous rehmannia, this method comprises the step of：Step 1：The processing of wine glutinous rehmannia；The preparation of step 2, test solution：The preparation of step 3, reference substance solution；Step 4, the foundation of regression equation and step 5 assay.QIJU DIHUANG KOUFUYE multicomponent quantitative analysis method provided by the present invention objectively can characterize the quality of QIJU DIHUANG KOUFUYE comprehensively.The screening technique of the processing procedure of wine glutinous rehmannia provided by the invention, using the active constituent of prepared rehmannia root as index, and best Preparation process method is filtered out by preferred content assaying method, entire process, operability is strong, scientific basis can be provided for the Preparation process of prepared rehmannia root, to controlling the quality of prepared rehmannia root and ensureing that clinical efficacy is of great significance.
Description
Technical field
The present invention relates to a kind of concocting methods of Chinese medicine, and in particular to the screening technique of the processing procedure of wine glutinous rehmannia.
Background technology
Glutinous rehmannia is the fresh or dried root of scrophulariaceae rehmannia glutinosa plant Rehmannia glutinosa Libosch., Radix Rehmanniae
Pharmacological property is changed into warm nature by cold after processing maturely, acts on by being converted into benefit clearly, taste turns sweet tea by hardship.Radix rehmanniae recen has heat-clearing cool
Blood, the effect of nourishing Yin and promoting production of body fluid, prepared rehmannia root then have the function of taking juice method, the Northern and Southern Dynasties to have steaming roasting, Sui after nourishing yin and supplementing blood, Han dynasty have steaming
Tang have in period wine mix steaming, steam quick-fried nine times, it is the methods of parched with wine, the Song Dynasty, which has, fries charcoal, vinegar-fried, ginger with the methods such as frying.Wine system is prepared rehmannia root
One of traditional concocting method, recording the when of processing according to ancient method requires to reach " nine, which steam nine, shines ", can just access have good quality it is ripe
Ground medicinal material.The prepared rehmannia root of commercial type has change more in Processing methods technique according to incompletely statistics now, especially auxiliary
In the use of material and on concocted time, and it can all simplify technique mostly.Majority is only with general characters such as " black as painted, sweet tea such as maltosemalt sugar "
Become the standard for being turned to reach processing end-point, there is no objective quality inspection standards.
Different time node of the present invention in prepared rehmannia root " nine, which steam nine, shines " concocting process is sampled research, using HPLC
Technology carries out containing measurement four kinds of Catalpol, acteoside, martynoside D and motherwort glycosides active constituents containing in prepared rehmannia root
It is fixed, preferably go out the processing procedure of best wine glutinous rehmannia.
Invention content
Goal of the invention：Present invention aims to solve the deficiencies of the prior art, and provides a kind of a kind of processing procedures of wine glutinous rehmannia
Screening technique should provide scientific basis for the Preparation process of prepared rehmannia root.To controlling the quality of prepared rehmannia root and ensureing clinical efficacy tool
It is significant.
Technical solution：To achieve the goals above, the technical solution that the present invention takes is：
A kind of screening technique of the processing procedure of wine glutinous rehmannia, includes the following steps：
Step 1：The processing of wine glutinous rehmannia
Take radix rehmanniae recen medicinal material, first day clean appearance silt dries spare, weighs yellow rice wine and mixes thoroughly, the bored profit of tinning sealing 24
Hour；Second day, medicinal material is put into Steam by water bath in pot；Medicinal material is placed in daylight decentralization drying and done, completed a steaming one and shine by third day
Concocting process；So again repeatedly eight times, nine overall processes for steaming the processing that nine shine are completed altogether；In concocting process, steam each time
It shines and leaves and takes part prepared rehmannia root and give over to sample for analysis；
Step 2：The preparation of test solution
It takes nine times and steams the prepared rehmannia root sample shone after processing, be respectively placed in baking oven dry, cooling, play powder sieving；Precision claims
Prepared rehmannia root coarse powder is taken, is placed in conical flask with cover, 50% methanol is added, takes out and is put to room temperature after supersound process, it is again weighed heavy
Amount, supplies weightlessness with 50% methanol, takes supernatant, centrifuges, after being processed to get nine steaming solarizations with 0.45 μm of miillpore filter filtration
Prepared rehmannia root test solution is saved backup in 4 DEG C；
Step 3：The preparation of reference substance solution
Take acteoside, Catalpol, martynoside D and motherwort glycosides reference substance appropriate respectively, it is accurately weighed, it is placed in volumetric flask
In, add methanol to be made into reference substance storing solution, it is spare to set 4 DEG C of refrigerators；The accurate above-mentioned reference substance solution of absorption adds methanol in right amount respectively
The mixed reference substance solution of acteoside, Catalpol, martynoside D and motherwort glycosides is made in constant volume；
Step 4：The foundation of regression equation
2,5,10,15,20, the 25 μ L of mixed reference substance solution of accurate aspiration step 3 inject liquid chromatograph, record respectively
Peak area carries out linear regression to sample size X with chromatographic peak area Y, has as a result shown good linear relationship；
Step 5：Assay
Nine steamings of accurate absorption shine 10 μ L of the prepared rehmannia root test solution after processing and inject liquid chromatograph, record peak respectively
Area, it is female according to acteoside, Catalpol, martynoside D and the benefit in the regression equation calculation prepared rehmannia root test solution of step 4
The content of careless glycosides.
Preferably, the screening technique of the processing procedure of above-described wine glutinous rehmannia,
Step 1：
It by 3 crowdes of separately sampled 300g of medicinal material, is numbered within first day, and cleans appearance silt, dry spare.After weigh
150g yellow rice wine is mixed thoroughly, the bored profit of tinning sealing 24 hours.Second day, 3 batches of medicinal materials are entered into pot Steam by water bath 8 hours respectively, water temperature is set
It is 100 DEG C.3 batches of medicinal materials are placed in daylight decentralization drying and done by third day, as the concocting process of " one, which steams one, shines ".It is so anti-
It is eight times multiple, nine overall processes for steaming the processing that nine shine are completed altogether；In concocting process, steaming solarization each time is left and taken part prepared rehmannia root and is given over to
For the sample of analysis.
Preferably, the screening technique of the processing procedure of above-described wine glutinous rehmannia,
Step 2：The preparation of test solution
It takes nine times and steams each 50g of prepared rehmannia root sample shone after processing, be placed in drying 8 hours in 80 DEG C of baking ovens, be put into drier
Interior cooling, beat powder cross 24 mesh sieve it is spare；Precision weighs coarse powder 2.0g, is placed in 50mL conical flask with cover, and accurate measurement is added
50% methanol 25ml is ultrasonically treated 1h, power 300W, frequency 40Hz, and taking-up is put to room temperature, again weighed weight, with 50% first
Alcohol supplies weightlessness, and supernatant 5mL, 8000rpm is taken to centrifuge 10min, is steamed to get nine times with 0.45 μm of miillpore filter filtration and shines processing
Prepared rehmannia root test solution afterwards, saves backup in 4 DEG C.
Preferably, the screening technique of the processing procedure of above-described wine glutinous rehmannia, step 3：Reference substance solution
It prepares
Take acteoside, Catalpol, martynoside D and motherwort glycosides reference substance appropriate respectively, it is accurately weighed, it is placed in 10mL appearances
In measuring bottle, methanol is added to be made into the reference substance storing solution that concentration is respectively 498 μ g/mL, 494 μ g/mL, 486 μ g/mL, 650 μ g/mL,
It is spare to set 4 DEG C of refrigerators；The accurate above-mentioned reference substance solution of absorption adds methanol constant volume to 10mL in right amount respectively, and every 1mL stamens containing hair are made
Spend glucosides, Catalpol, martynoside D and motherwort glycosides molten for 49.80 μ g, 98.80 μ g, 97.20 μ g, the mixing reference substance of 130.0 μ g
Liquid.
Preferably, the screening technique of the processing procedure of above-described wine glutinous rehmannia, the chromatography of step 4 and step 5
Condition is：
Kromasil 100-5C18 chromatographic columns, 4.6mm × 250mm, 5 μm, mobile phase acetonitrile and 0.1% phosphoric acid water；Gradient
Elution：0~5min, 2%A；5~20min, 20%A；20~25min, 30%A；25min~40min, 30%A；40~
45min, 2%A；45~50min, 2%A, Detection wavelength 203/334nm, 35 DEG C of column temperature, 20 μ L of sample size.
Preferably, the screening technique of the processing procedure of above-described wine glutinous rehmannia, the verbascose that step 4 is established
Glycosides, Catalpol, martynoside D and motherwort glycosides regression equation are：
Craft screening is tested：
The present invention is unfolded to study ancient method processing prepared rehmannia root in the chemical composition of different concocted time nodes for the first time, right
Contain quantitative change in " nine steam nine shine " concocting process with four kinds of acteoside, Catalpol, martynoside D and motherwort glycosides active constituents
The comparative study for changing the more system that carried out, scientific basis is provided for glutinous rehmannia concocted technology.
In the separation feelings for investigating acteoside, Catalpol, martynoside D and motherwort glycosides four kinds of tested ingredients and impurity peaks
When condition, three kinds of -0.1% phosphate aqueous solution of acetonitrile, -0.1% formic acid water of acetonitrile, acetonitrile-water flowings are first compared in preliminary experiment
As a result the gradient elution effect of phase is shown so that -0.1% phosphoric acid water of acetonitrile is preferable as mobile phase peak shape, separating degree is high, baseline is flat
Surely, therefore -0.1% phosphoric acid water of final choice acetonitrile is mobile phase.
Screening experiment of the present invention finds four kinds of acteoside, Catalpol, martynoside D and motherwort glycosides index ingredients most
Big absorbing wavelength is respectively 334nm, 203nm, 203nm and 210nm, is finding acteoside content in wave in preliminary experiment
It is relatively low at long 203nm, and there is notable difference under maximum absorption wavelength 334nm, and Catalpol, martynoside D and motherwort glycosides are in 334nm
Almost without being absorbed under wavelength, motherwort glycosides content does not have notable difference at maximum absorption wavelength 210nm and 203nm, because
This selects 203nm as the Detection wavelength of three kinds of Catalpol, martynoside D and motherwort glycosides ingredients, and 334nm is as acteoside
Detection wavelength.
The present invention in sample pre-treatments, respectively to extracting mode (ultrasound, be heated to reflux), Extraction solvent (25% methanol,
50% methanol, 100% methanol, acetonitrile), extraction time (30,60,90min), sampling amount (0.5,1.0,2.0g) examined
It examines, the results showed that test solution repeatability made of being heated to reflux>3%, preliminary error in judgement comes across sentencing for the degree of being evaporated
It is fixed, cause part test solution concentration larger, part test solution concentration is just smaller, so method can not be taken.Due to
Ultrasound procedure is easy, and feasibility is high, therefore takes the method extraction of ultrasound.
When being investigated to different solvents, the methanol extraction content difference of various concentration is small, and is extracted with acetonitrile
When there are more impurity peaks, finally select a concentration of 50% methanol as Extraction solvent.
When being investigated to extraction time, extracts chemical composition at duration 30min and extract incomplete, extraction time
60min and 90min differences are smaller, therefore a length of 60min when selective extraction.
When being investigated to sampling amount, four kinds of index ingredient responses when sampling amount is 0.5g and 1.0g are as a result shown
It is relatively low, therefore select sampling amount for 2.0g.
By the above screening experiment, 50% methanol 25mL is added in final choice 2.0g sample powders, is ultrasonically treated 60min
The method prepared for this experiment test solution.
Advantageous effect：
1, the screening technique of the processing procedure of wine glutinous rehmannia provided by the invention, using the active constituent of prepared rehmannia root as index, and
Best Preparation process method is filtered out by preferred content assaying method, entire process, operability is strong, can be ripe
The Preparation process of glutinous rehmannia provides scientific basis, to controlling the quality of prepared rehmannia root and ensureing that clinical efficacy is of great significance.
2, the screening technique of the processing procedure of wine glutinous rehmannia provided by the invention, with method is easy, stability is good, precision
High, high repeatability and other advantages.
Description of the drawings
Fig. 1 is chromatogram of the mixed reference substance solution at 203nm.
Fig. 2 is chromatogram of the wine glutinous rehmannia test sample at 203nm.
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.
Following embodiment instrument
Japanese Shimadzu Corporation Shimadzu LC-20AB highly effective liquid phase chromatographic systems, including on-line degassing machine, automatic sampling
Device Prominence SIL-20A, diode array detector SPD-M20A and column oven CTO-20A；ME204E electronic analysis day
Flat (plum Teller-support benefit Internaional, Inc, be accurate to ten a ten thousandths)；BSM220.4 assay balances (the tall and erect essence in Shanghai
Company is accurate to a ten thousandth)；(1) Ying Heng companies are accurate to percent to square plate electronic scale；DSX-18L high-pressure sterilizing pots (on
Hai Shenan equipment Co., Ltd)；GVS types ultrasonic cleaner (Gou Wei Science and Technology Ltd.s of Shenzhen)；DY-20 continuous-flow type Chinese medicines
Pulverizer (power Chinese medicine instrument Co., Ltd of Austria of Wenling city)；(upper Nereid is macro for DHG-9140A type vertical electrics constant temperature blast drying oven
Equipment Co., Ltd).
Reagent used in following embodiment
Acteoside reference substance (lot number 61276-17-3, purity >=98%), motherwort glycosides reference substance (lot number 52949-
83-4, purity >=98%), martynoside D reference substances (lot number 81720-08-3, purity >=98%), Catalpol reference substance (lot number
2415-24-9, purity >=98%) it is purchased from the Nanjing bio tech ltd Sen Beijia.Three batches of glutinous rehmannia samples are purchased from river respectively
Nan Sheng Yong Xintang prepared slices of Chinese crude drugs Science and Technology Ltd., Anhui Hu Qiao Chinese medicines Co., Ltd and the Zhejiang Tong Juntang prepared slices of Chinese crude drugs are limited
Company.It is accredited as scrophulariaceae rehmannia glutinosa plant (Rehmannia through Nanjing Hai Yuan prepared slices of Chinese crude drugs Co., Ltd pharmacist of traditional Chinese medicine's fourth is striking
Glutinosa Libosch) dry rhizome.Acetonitrile, methanol are chromatographically pure, and water is ultra-pure water, and phosphoric acid is that analysis is pure.GB/T
13662 yellow rice wine (are purchased from Guyuelongshan Shaoxing Wine Co Ltd, Zhejiang).
1, a kind of screening technique of the processing procedure of wine glutinous rehmannia, includes the following steps：
Step 1：The processing of wine glutinous rehmannia
It by 3 crowdes of separately sampled 300g of medicinal material, is numbered within first day, and cleans appearance silt, dry spare.After weigh
150g yellow rice wine is mixed thoroughly, the bored profit of tinning sealing 24 hours.Second day, 3 batches of medicinal materials are entered into pot Steam by water bath 8 hours respectively, water temperature is set
It is 100 DEG C.3 batches of medicinal materials are placed in daylight decentralization drying and done by third day, as the concocting process of " one, which steams one, shines ".It is so anti-
It is eight times multiple, nine overall processes for steaming the processing that nine shine are completed altogether；In concocting process, steaming solarization each time is left and taken part prepared rehmannia root and is given over to
For the sample of analysis；
Step 2：The preparation of test solution
It takes nine times and steams each 50g of prepared rehmannia root sample shone after processing, be placed in drying 8 hours in 80 DEG C of baking ovens, be put into drier
Interior cooling, beat powder cross 24 mesh sieve it is spare；Precision weighs coarse powder 2.0g, is placed in 50mL conical flask with cover, and accurate measurement is added
50% methanol 25ml is ultrasonically treated 1h, power 300W, frequency 40Hz, and taking-up is put to room temperature, again weighed weight, with 50% first
Alcohol supplies weightlessness, and supernatant 5mL, 8000rpm is taken to centrifuge 10min, is steamed to get nine times with 0.45 μm of miillpore filter filtration and shines processing
Prepared rehmannia root test solution afterwards, saves backup in 4 DEG C；
Step 3：The preparation of reference substance solution
Take acteoside, Catalpol, martynoside D and motherwort glycosides reference substance appropriate respectively, it is accurately weighed, it is placed in 10mL appearances
In measuring bottle, methanol is added to be made into the reference substance storing solution that concentration is respectively 498 μ g/mL, 494 μ g/mL, 486 μ g/mL, 650 μ g/mL,
It is spare to set 4 DEG C of refrigerators；The accurate above-mentioned reference substance solution of absorption adds methanol constant volume to 10mL in right amount respectively, and every 1mL stamens containing hair are made
Spend glucosides, Catalpol, martynoside D and motherwort glycosides molten for 49.80 μ g, 98.80 μ g, 97.20 μ g, the mixing reference substance of 130.0 μ g
Liquid；
Step 4：The foundation of regression equation
2,5,10,15,20, the 25 μ L of mixed reference substance solution of accurate aspiration step 3 inject liquid chromatograph, record respectively
Peak area, such as Fig. 1 carry out linear regression to sample size X with chromatographic peak area Y, have as a result shown good linear relationship；
Step 5：Assay
Nine steamings of accurate absorption shine 10 μ L of the prepared rehmannia root test solution after processing and inject liquid chromatograph, record peak respectively
Area, such as Fig. 2, according to acteoside, Catalpol, the martynoside D in the regression equation calculation prepared rehmannia root test solution of step 4
With the content of motherwort glycosides.
The chromatographic condition of the screening technique of the processing procedure of above-described wine glutinous rehmannia, step 4 and step 5 is：
Kromasil 100-5C18 chromatographic columns, 4.6mm × 250mm, 5 μm, mobile phase acetonitrile and 0.1% phosphoric acid water；Gradient
Elution：0~5min, 2%A；5~20min, 20%A；20~25min, 30%A；25min~40min, 30%A；40~
45min, 2%A；45~50min, 2%A, Detection wavelength 203/334nm, 35 DEG C of column temperature, 20 μ L of sample size.
Acteoside, Catalpol, martynoside D and motherwort glycosides regression equation such as the following table 1 that step 4 is established：
1 regression equation of table
2 methodological study of embodiment
1, precision test
Precision draws the mixed reference substance solution of above-described embodiment 1, by the chromatographic condition of above-described embodiment 1, continuous sample introduction 6
It is secondary, record peak area.It is 0.27% to measure acteoside peak area RSD, and Catalpol peak area RSD is 0.55%, the peaks martynoside D
Area RSD is 1.07%, and motherwort glycosides peak area RSD is 0.77%.Four kinds of index ingredient reference substance solution RSD values are respectively less than
3%, the results showed that this test apparatus precision is good.
2, repeatability is investigated
Precision is weighed with a collection of 6 parts of prepared rehmannia root sample powder, prepares sample solution by the method for embodiment step 2, respectively
It measures, records chromatogram, as a result show that acteoside RSD is 1.29%, Catalpol RSD is 1.15%, and martynoside D RSD are
1.88%, motherwort glycosides RSD are 2.44%.Each sample RSD values are respectively less than 3%, the results showed that this test method repeatability is good.
3, stability test
Precision is drawn with a test solution, by the chromatographic condition of above-described embodiment 1 respectively at 0,2,4,8,12, for 24 hours
Sample introduction, the peak area for recording four ingredients accumulate peak value, as a result show that acteoside RSD is 2.56%, Catalpol RSD is
0.95%, martynoside D RSD are 1.78%, and motherwort glycosides RSD is 2.48%.Each sample RSD values are respectively less than 3%, the results showed that
Test solution is good in internal stability for 24 hours.
4, sample recovery rate is tested
Precision is weighed from 6 parts of sample powder 2.0g of a collection of prepared rehmannia root respectively, is respectively placed in conical flask, precision is added
Suitable reference substance so that contain 300.0 μ g of acteoside, Catalpol 600.0 μ g, martynoside D in sample solution respectively
900.0 μ g, 1000 μ g of motherwort glycosides.According to 3.2.2 lower section legal system available test sample solutions, by the chromatostrip of above-described embodiment 1
Part measures, and calculates the average recovery rate and RSD of each ingredient, shows that stability is good, be specifically shown in Table 2.
The sample recovery rate experiment of four index ingredients in 2 prepared rehmannia root of table
Embodiment 3
1, sample measures
It fetches from 3 batches of different sources and processes the prepared rehmannia root powder completed, weigh powder 2g respectively, it is accurately weighed, by real
It applies 1 time processing method of example and prepares sample solution, and measured by 1 time chromatographic condition of embodiment, calculate content and be shown in Table 3, table 4, table 5.
Table 3 prepared rehmannia root medicinal material (Henan) assay (mgg-1)
Table 4 prepared rehmannia root medicinal material (Anhui) assay (mgg-1)
Table 5 prepared rehmannia root medicinal material (Zhejiang) assay (mgg-1)
The experimental results showed that, Catalpol, martynoside D and motherwort glycosides content are presented with the increase for steaming number above
Different degrees of decline.Tentatively judge to belong to iridoid glycosides compound because of Catalpol, martynoside D and motherwort glycosides, and
The universal polarity of iridoid is larger, and soluble easily in water and thermal stability is poor.Its palliating degradation degree master during heating
Related to the number of sugar, three glucosides are hardly degraded, bioside degradation part, and monoglycosides almost all is degraded.Consider because
Catalpol and motherwort glycosides are monoglycosides, and thermal stability is poor, thus both ingredients with steam number increase, content be decreased obviously and
Amplitude is larger.Though martynoside D contents have decline with steaming number increase, tend towards stability after " five steam five shine ".Feltwort
Glucosides belongs to benzyl carbinol glycoside compound, the results showed that its content at " six steam six shine " reaches maximum value, with steaming number
Increase after content declined.
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 (5)
1. a kind of screening technique of the processing procedure of wine glutinous rehmannia, which is characterized in that include the following steps：
Step 1：The processing of wine glutinous rehmannia
Take radix rehmanniae recen medicinal material, first day clean appearance silt dries spare, weighs yellow rice wine and mixes thoroughly, the bored profit of tinning sealing 24 hours；
Second day, medicinal material is put into Steam by water bath in pot；Medicinal material is placed in the big gun that daylight decentralization drying is dry, and one steaming one of completion is shone by third day
Process processed；So again repeatedly eight times, nine overall processes for steaming the processing that nine shine are completed altogether；In concocting process, steams to shine each time and stay
Part prepared rehmannia root is taken to give over to the sample for analysis；
Step 2：The preparation of test solution
It takes nine times and steams the prepared rehmannia root sample shone after processing, be respectively placed in baking oven dry, cooling, play powder sieving；Precision weighs ripe
Glutinous rehmannia coarse powder, is placed in conical flask with cover, and 50% methanol is added, and takes out and is put to room temperature after supersound process, weighed weight, is used again
50% methanol supplies weightlessness, takes supernatant, centrifugation, steams the radix rehmanniae preparata shone after processing with 0.45 μm of miillpore filter filtration to get nine times
Yellow test solution is saved backup in 4 DEG C；
Step 3：The preparation of reference substance solution
Take acteoside, Catalpol, martynoside D and motherwort glycosides reference substance appropriate respectively, it is accurately weighed, it is placed in volumetric flask,
Add methanol to be made into reference substance storing solution, it is spare to set 4 DEG C of refrigerators；The accurate above-mentioned reference substance solution of absorption adds methanol fixed in right amount respectively
Hold, the mixed reference substance solution of acteoside, Catalpol, martynoside D and motherwort glycosides is made；
Step 4：The foundation of regression equation
2,5,10,15,20, the 25 μ L of mixed reference substance solution of accurate aspiration step 3 inject liquid chromatograph respectively, record peak face
Product carries out linear regression to sample size X with chromatographic peak area Y, has as a result shown good linear relationship；
Step 5：Assay
Nine steamings of accurate absorption shine 10 μ L of the prepared rehmannia root test solution after processing and inject liquid chromatograph, record peak face respectively
Product, according to acteoside, Catalpol, martynoside D and the motherwort in the regression equation calculation prepared rehmannia root test solution of step 4
The content of glycosides.
2. the screening technique of the processing procedure of wine glutinous rehmannia according to claim 1, which is characterized in that step 2：Test sample is molten
The preparation of liquid
It takes nine times and steams each 50g of prepared rehmannia root sample shone after processing, be placed in drying 8 hours in 80 DEG C of baking ovens, be put into cold in drier
But, beat powder cross 24 mesh sieve it is spare；Precision weighs coarse powder 2.0g, is placed in 50mL conical flask with cover, and accurate 50% measured is added
Methanol 25ml is ultrasonically treated 1h, power 300W, frequency 40Hz, and taking-up is put to room temperature, and weighed weight, is mended with 50% methanol again
Foot weightlessness takes supernatant 5mL, 8000rpm to centrifuge 10min, after shining processing with 0.45 μm of miillpore filter filtration to get nine steamings
Prepared rehmannia root test solution is saved backup in 4 DEG C.
3. the screening technique of the processing procedure of wine glutinous rehmannia according to claim 1, which is characterized in that step 3：Reference substance is molten
The preparation of liquid
Take acteoside, Catalpol, martynoside D and motherwort glycosides reference substance appropriate respectively, it is accurately weighed, it is placed in 10mL volumetric flasks
In, add methanol to be made into the reference substance storing solution that concentration is respectively 498 μ g/mL, 494 μ g/mL, 486 μ g/mL, 650 μ g/mL, sets 4
DEG C refrigerator is spare；The accurate above-mentioned reference substance solution of absorption adds methanol constant volume to 10mL in right amount respectively, and every 1mL is made and contains verbascose
Glycosides, Catalpol, martynoside D and motherwort glycosides are the mixed reference substance solution of 49.80 μ g, 98.80 μ g, 97.20 μ g, 130.0 μ g.
4. the screening technique of the processing procedure of wine glutinous rehmannia according to claim 1, which is characterized in that step 4 and step 5
Chromatographic condition is：
Kromasil 100-5C18 chromatographic columns, 4.6mm × 250mm, 5 μm, mobile phase acetonitrile and 0.1% phosphoric acid water；Gradient is washed
It is de-：0~5min, 2%A；5~20min, 20%A；20~25min, 30%A；25min~40min, 30%A；40~45min,
2%A；45~50min, 2%A, Detection wavelength 203/334nm, 35 DEG C of column temperature, 20 μ L of sample size.
5. the screening technique of the processing procedure of wine glutinous rehmannia according to claim 1, which is characterized in that the hair that step 4 is established
Stamen flower glucosides, Catalpol, martynoside D and motherwort glycosides regression equation are：
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2168588A1 (en) * | 2007-07-18 | 2010-03-31 | Ling Zhang | An extract of rehmannia glutinasa libosch. for reducing blood sugar, reducing blood fat, treating leukemia and preparation method and uses thereof |
| CN102072941A (en) * | 2010-11-19 | 2011-05-25 | 四川美大康药业股份有限公司 | Fingerprint test method for adhesive rehmannia leaf total glycoside |
-
2018
- 2018-06-21 CN CN201810641930.3A patent/CN108802229A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2168588A1 (en) * | 2007-07-18 | 2010-03-31 | Ling Zhang | An extract of rehmannia glutinasa libosch. for reducing blood sugar, reducing blood fat, treating leukemia and preparation method and uses thereof |
| CN102072941A (en) * | 2010-11-19 | 2011-05-25 | 四川美大康药业股份有限公司 | Fingerprint test method for adhesive rehmannia leaf total glycoside |
Non-Patent Citations (3)
| Title |
|---|
| 卢鹏伟: "地黄的化学成分和炮制的比较研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 孟祥龙 等: "熟地黄炮制（九蒸九晒）过程中药效化学成分量变化及炮制辅料对其影响研究", 《中草药》 * |
| 张留记 等: "HPLC同时测定地黄中5中苷类成分的含量", 《天然产物研究与开发》 * |
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Application publication date: 20181113 |