CN108037234A - A kind of quality determining method of Guanule contg. ' Jigucao particle - Google Patents
A kind of quality determining method of Guanule contg. ' Jigucao particle Download PDFInfo
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Abstract
The invention discloses a kind of quality determining method of Guanule contg. ' Jigucao particle, including the content detection of thin-layer chromatography Qualitive test and aurantiamarin.The present invention establishes the thin-layer chromatography Qualitive test of hypericum japonicum in Guanule contg. ' Jigucao particle first, and assay is carried out to aurantiamarin, the detection method has stronger specificity, durability, its accuracy, reappearance, stability can reach the requirement of research and production, so as to which the stability of product quality and controllability is effectively ensured.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to a kind of quality determining method of Guanule contg. ' Jigucao particle.
Background technology
Guanule contg. ' Jigucao particle is made of Canton love-pea vine, oriental wormwood, hypericum japonicum, downy rosemyrtle root, ghostplant wormwood herb, 6 taste Chinese medicine of avicenna pricklyash root,
Record in Ministry of Health of the People's Republic of China's standard Traditional Chinese medicine historical preparation the 3rd copy page 94, standard No.：WS3-B-0561-91,
For oxyhepatitis icteric or non-icteric type good effect, removing jaundice is fast, can remove virus and protect liver, effectively reduce hepatitis
The sequelae of patient.The quality standard of Guanule contg. ' Jigucao particle only has character, granule inspection item and microbial limit inspection
Project is tested, but there is no thin layer to differentiate that item and assay item, quality standard controllability are too low, it is difficult to monitors the inherent matter of product
Amount, method of quality control have much room for improvement.
Journey waits document report quietly《The thin layer of Guanule contg. ' Jigucao particle differentiates and chlorogenic acid, the measure of Quercetin》Using
TLC carries out Qualitive test to Canton love-pea vine.There are the following problems for this method：1., this method use 366nm ultraviolet lamps under inspect,
366nm wavelength is of little use in thin-layer identification method,《Chinese Pharmacopoeia》In middle 0502 thin-layered chromatography of general rule of version four in 2015
Checking device has visible ray, 254nm and 365nm ultraviolet sources, and many laboratories do not possess 366nm ultraviolet lamps, illustrate the party
Method is not easy to promote and apply.2., in addition, thin layer figure principal spot Rf values are bigger than normal in the discrimination method, principal spot has gone to solvent front,
Do not meet《Chinese Pharmacopoeia》(Rf to be between 0.2~0.8 for the middle 0502 thin-layered chromatography Rf of general rule of version four in 2015 requirements
Preferably).3., the discrimination method do not carry out serviceability test, it is impossible to ensure that established method can be used for routine check and side
The reliability of method.
Have document report using HPLC measure at the same time Guanule contg. ' Jigucao particle Content of Chlorogenic Acid, Quercetin and protocatechualdehyde,
The content of protocatechuic acid, gallic acid.It is well known, however, that Quercetin, protocatechualdehyde, protocatechuic acid, gallic acid etc. are extensive
In the presence of with medicinal plant, specificity is lacked using it as index components.
In addition, journey waits document report quietly《The thin layer of Guanule contg. ' Jigucao particle differentiates and chlorogenic acid, the measure of Quercetin》
The content of middle measure chlorogenic acid, the active ingredient of oriental wormwood in the chlorogenic acid side of being attributed to.It is worth noting that,《Chinese Pharmacopoeia》2015
Regulation is different according to collecting season in year one, and the spring habit of harvesting claims " capillary wormwood ", and the autumn habit of harvesting claims " spending oriental wormwood ".It is green
Ortho acid is capillary wormwood assay index, cannot be less than 0.50%；Escoparone is flower oriental wormwood assay index, be cannot be less than
0.20%.But do not indicate which season harvesting oriental wormwood is in Guanule contg. ' Jigucao particle prescription.There is scholar to report capillary wormwood
The content of Content of Chlorogenic Acid is 0.81%, and the content of flower oriental wormwood Content of Chlorogenic Acid is 0.46%, and capillary wormwood is than the chlorogenic acid in colored oriental wormwood
It is high.Content's index component of this explanation by the use of chlorogenic acid as Guanule contg. ' Jigucao particle is unreasonable, it is impossible to by measuring Canton love-pea vine
The rate of transform of Ganyan Granule Content of Chlorogenic Acid height dereaction index components and the controllability for evaluating product quality.
The document reports such as Hou little Tao《Hplc simultaneous determination Guanule contg. ' Jigucao particle protocatechuic acid, original
The content of catechu aldehyde and gallic acid》In prepare test solution during, ultrasound carries after sample needs low temperature drying to constant weight
Take, there are the shortcomings that cumbersome, the consuming time is long.
The content of the invention
In order to overcome the above-mentioned deficiencies of the prior art, it is an object of the invention to provide a kind of matter of Guanule contg. ' Jigucao particle
Quantity measuring method, this method specificity is good, durability is strong, and stability is good, can effectively ensure that product quality stability and can
Control property.
The present invention is to be achieved through the following technical solutions：
A kind of quality determining method of Guanule contg. ' Jigucao particle, it is characterised in that including thin-layer chromatography Qualitive test and orange
The content detection of skin glycosides：
(1) the thin-layer chromatography Qualitive test of hypericum japonicum：
This product 3g is taken, it is finely ground, add water 25ml, 20-40 minutes ultrasonic, filtration, adds dilute hydrochloric acid tune pH 1~2, with acetic acid second
Ester extracts 1-3 times, and each 20ml, combined ethyl acetate extract, is evaporated, and residue adds ethanol 1ml to make dissolving, molten as test sample
Liquid；Hypericum japonicum control medicinal material 1g separately is taken, adds water 25ml, is heated to reflux 20-40 minutes, is filtered, " adds dilute hydrochloric acid tune pH 1~2 " certainly
Rise and be made in the same way of hypericum japonicum control medicinal material solution, it is spare；Tested according to thin-layered chromatography, draw each 5~10 μ l of above two solution,
Put respectively on same silica gel g thin-layer plate, with volume ratio dichloromethane：Ethyl acetate：Methanol：Formic acid：Water is 12:10:2:1:1
For solvent, it is unfolded, takes out, dry, spray with alchlor test solution, heats 5~10min at 105 DEG C, put under 365nm ultraviolet lamps
Inspect；In test sample chromatography, on position corresponding with control medicinal material chromatography, same color spot；
(2) assay of aurantiamarin：
It is measured using high performance liquid chromatography, specific method and step are as follows：
Chromatographic condition is filler with octadecylsilane chemically bonded silica with system suitability；With volume ratio acetonitrile-
Water 17：83 be mobile phase；Detection wavelength is 283nm；Number of theoretical plate is calculated by aurantiamarin peak should be not less than 5000；
The preparation of reference substance solution takes aurantiamarin reference substance appropriate, accurately weighed, adds methanol that every 1ml is made molten containing 15 μ g
Liquid, to obtain the final product；
The preparation of test solution takes this product, finely ground, crosses No. five sieves, takes 2.0g, accurately weighed, puts in conical flask with cover,
Precision adds 10-50% methanol 50ml, and weighed weight, is ultrasonically treated 30-60 minutes, takes out, lets cool, then weighed weight, uses 10-
50% methanol supplies the weight of less loss, shakes up, and filtration, takes subsequent filtrate, to obtain the final product；
Determination method difference is accurate to draw reference substance solution and each 10 μ l of test solution, injects liquid chromatograph, measures, i.e.,
.
Preferably, in described (3), test solution is prepared as：This product is taken, it is finely ground, No. five sieves are crossed, take 2.0g, it is accurate
It is weighed, to put in conical flask with cover, precision adds 10% methanol 50ml, and weighed weight, is ultrasonically treated 45 minutes, takes out, lets cool, then
Weighed weight, the weight of less loss is supplied with 10% methanol, is shaken up, and filtration, takes subsequent filtrate, to obtain the final product.
The present invention carries out thin-layer qualitative discriminating to hypericum japonicum in Guanule contg. ' Jigucao particle, can be inspected under 365nm ultraviolet lamps,
Specificity is strong, and Rf values are moderate, and about 0.2~0.5, and two principal spots have good separating degree (R>1.5), and this method has
Durability, it is ensured that the reliability of method, suitable for promoting and applying.
The present invention selects aurantiamarin as assay index components, index components and work of the aurantiamarin for Chinese medicine avicenna pricklyash root
Property component, and avicenna pricklyash root be prescription in dosage maximum medicinal material.Canton love-pea vine is monarch drug in a prescription in side, according to《Chinese Pharmacopoeia》2015
One its quality standard of version is without assay item.Further consulting literatures,《Hong Kong Chinese medicine》6th phase recorded Canton love-pea vine medicinal material,
The content limit of abrine is calculated not less than 0.025% by dry product in regulation Canton love-pea vine.Since abrine is slightly soluble in water,
And the technique of Guanule contg. ' Jigucao particle uses water decocting herbs, the recovery rate that this directly results in abrine is low.Therefore, we taste
The content of pilot production HPLC detection abrines, result of the test show that its content is extremely low, should not measure its content.So this hair
The content of aurantiamarin is rational in bright measure Guanule contg. ' Jigucao particle, and aurantiamarin has certain specificity, physics, chemistry
Property is stablized, and reduces the introducing of systematic error, improves the accuracy and reappearance of measurement result, ensures Guanule contg. ' Jigucao particle
The stability and quality controllability of quality.
The present invention prepares test solution using direct ultrasonic extracting method, and ultrasound carries after eliminating low temperature drying to constant weight
Take the operation of measure.This method is simple, convenient, time saving, environmentally friendly, avoids causing index components need not because operating process is cumbersome
The loss wanted, at utmost reflects the real content of aurantiamarin index components.
Compared with prior art, the present invention have the advantages that：
The present invention establishes the thin-layer chromatography Qualitive test of hypericum japonicum in Guanule contg. ' Jigucao particle first, and to aurantiamarin into
Assay is gone, which has stronger specificity, durability, its accuracy, reappearance, stability can reach
The requirement of research and production, can effectively ensure that the stability and controllability of product quality.
Brief description of the drawings
Fig. 1 is that the thin layer of hypericum japonicum differentiates chromatogram；Wherein 1. it is the negative sample of hypericum japonicum, 2. -4. it is Canton love-pea vine liver
Scorching particulate samples, are 5. hypericum japonicum control medicinal material；
Fig. 2 is that the thin layer of hypericum japonicum in 10 batches of test samples differentiates chromatogram；Wherein 1. to lack the negative sample of hypericum japonicum,For 10 batches of Guanule contg. ' Jigucao particulate samples,For hypericum japonicum control medicinal material；
Fig. 3 is aurantiamarin reference substance canonical plotting；
Fig. 4 schemes for aurantiamarin reference substance HPLC；
Fig. 5 is to lack avicenna pricklyash root negative controls HPLC figures；
Fig. 6 schemes for Guanule contg. ' Jigucao particulate samples HPLC.
Embodiment
The present invention is further illustrated below by embodiment, and following embodiments are the specific embodiment party of the present invention
Formula, but embodiments of the present invention and from the limitation of following embodiments.
Embodiment 1：
A kind of quality determining method of Guanule contg. ' Jigucao particle, it is characterised in that including thin-layer chromatography Qualitive test and orange
The content detection of skin glycosides：
(1) the thin-layer chromatography Qualitive test of hypericum japonicum：
This product 3g is taken, it is finely ground, add water 25ml, 20-40 minutes ultrasonic, filtration, adds dilute hydrochloric acid tune pH 1~2, with acetic acid second
Ester extracts 1-3 times, and each 20ml, combined ethyl acetate extract, is evaporated, and residue adds ethanol 1ml to make dissolving, molten as test sample
Liquid；Hypericum japonicum control medicinal material 1g separately is taken, adds water 25ml, is heated to reflux 20-40 minutes, is filtered, " adds dilute hydrochloric acid tune pH 1~2 " certainly
Rise and be made in the same way of hypericum japonicum control medicinal material solution, it is spare；Tested according to thin-layered chromatography, draw each 5~10 μ l of above two solution,
Put respectively on same silica gel g thin-layer plate, with volume ratio dichloromethane：Ethyl acetate：Methanol：Formic acid：Water is 12:10:2:1:1
For solvent, it is unfolded, takes out, dry, spray with alchlor test solution, heats 5~10min at 105 DEG C, put under 365nm ultraviolet lamps
Inspect；In test sample chromatography, on position corresponding with control medicinal material chromatography, same color spot；
Methodology validation：
1st, specificity is investigated
In test sample chromatography, on position corresponding with hypericum japonicum control medicinal material chromatography, the spot of two same colors is shown.
And lack in hypericum japonicum negative sample chromatography, it is noiseless on a corresponding position.Illustrate discriminating tool of this method to hypericum japonicum medicinal material
There is specificity.The result is shown in Figure 1.
2nd, durability is investigated
The research of 2.1 different manufacturers lamellaes
Hand bed board (100 × 200mm, thickness 0.25mm), prefabricated board (the limited public affairs of Qingdao spectrum section separation material are investigated respectively
Department, 20161022,100 × 200mm, 0.20~0.25mm of thickness) and Merck plates (thickness 0.25mm), three kinds of thin layer chromatography boards
Influence to chromatographic behavior.The Rf values about 0.3~0.4 of hand bed board principal spot, the Rf values than prefabricated board, Merck plates are big, the above three
The lamellae of type can obtain good separating effect, illustrate that variation of this method to lamellae type has durability.
2.2 investigate influence of the expansion temperature to chromatographic behavior
Investigate 8 DEG C of different temperatures, 21 DEG C, 26 DEG C, 30 DEG C of influences to principal spot separating effect in collection of illustrative plates.The result is shown in
The principal spot Rf values that aforementioned four investigates collection of illustrative plates obtained by temperature are basically identical, do not produced with the change of temperature significant
Influence, illustrate that this method has durability to the variation that temperature is unfolded.Therefore, special rule are not made to expansion temperature in text
It is fixed.
2.3 investigate influence of the relative humidity to chromatographic behavior
It is 18%, 42%, 65%, 88% condition in relative humidity by each 10 μ l points of test solution in silica gel g thin-layer plate
Lower pre-equilibration 30min, expansion.The results show difference relative humidity influences little, equal energy to the principal spot of test solution chromatography
Good separating effect is obtained, illustrates that this method has durability to the variation that relative humidity is unfolded.Therefore, not to exhibition in text
Open relative humidity and make special provision.
2.4 investigate influence of the presaturation time to chromatographic behavior
Compare whether different presaturation time 0min, 15min, 30min make a significant impact thin-layer chromatography.The results show
The principal spot separation of each test solution chromatogram is all right, and it is durable to show that this method has the different presaturation time
Property.Therefore, special provision is not made to the presaturation time in text.
2.5 investigate influence of the different point sample amounts to chromatographic behavior
Investigate whether different 3 μ l of point sample amount, 5 μ l, 8 μ l, 10 μ l make a significant impact thin-layer chromatography.4 kinds of points more than
In the test sample chromatography of sample amount, principal spot can efficiently separate.During 3 μ l point sample amounts, since the few reason principal spot of point sample amount omits
It is aobvious fuzzy, but can recognize.When point sample amount is 5 μ l, 8 μ l, 10 μ l, thin-layer chromatogram effect is preferable, and principal spot is clearly easily distinguished.
Determine that point sample amount is 5~10 μ l in view of the content difference between different batches of product, in text.
2.6 investigate influence of the different solvents to chromatographic behavior
Investigate petroleum ether-ethyl acetate-methyl alcohol-formic acid=5 that volume ratio boiling range is 60~90 DEG C:3:1:0.2 expansion system
System, testing compound spot and R f value about 0.2, adjacent spots interfere compound spots to be measured, and separating degree does not reach requirement.
Investigation volume ratio is ethyl acetate-acetone-formic acid-water=5:3:0.5:05 development system, testing compound spot and R f value is about
0.9, adjacent spots interfere compound spots to be measured, and separating degree does not reach requirement.Investigation volume ratio is ethyl acetate-first
Alcohol-water-formic acid=100:1:1:2 development systems, testing compound spot and R f value about 0.5, adjacent spots are to testing compound spot
Point is noiseless, but spot trails.Investigation volume ratio is dichloromethane-ethyl acetate-methyl alcohol-formic acid-water=12:10:2:1:1 exhibition
Open system, testing compound spot and R f value about 0.5, in test sample chromatography, in position corresponding with hypericum japonicum control medicinal material chromatography
On, the spot of same color is shown, principal spot is clearly easily debated in thin layer figure, and separating degree is good, and adjacent spots are noiseless.Therefore select volume
Than for dichloromethane-ethyl acetate-methyl alcohol-formic acid-water=12:10:2:1:1 development system.
The inspection of 2.7 10 batches of test samples
The Guanule contg. ' Jigucao particle in 10 batches of different production periods is taken respectively, is prepared test solution and is tested, as a result
See Fig. 2.In 10 batches of test sample chromatographies, on position corresponding with control medicinal material chromatography, the spot of same color is shown.
To sum up, in the thin-layer chromatography Qualitive test of hypericum japonicum of the invention negative control on a corresponding position without dry
Disturb, show that this method specificity is strong；Rf values are moderate, and about 0.2~0.5, and two principal spots have good separating degree (R>
1.5)；Meanwhile different manufacturers lamellae, different temperatures, different relative humidity, different presaturation times, different point samples are investigated
There are no significant influences on hypericum japonicum control medicinal material chromatographic behavior for amount, different six kinds of factors of solvent, prompts this method to have resistance to
With property, it is ensured that the reliability of method, suitable for promoting and applying.
(2) assay of aurantiamarin：
It is measured using high performance liquid chromatography, specific method and step are as follows：
Chromatographic condition is filler with octadecylsilane chemically bonded silica with system suitability；With volume ratio acetonitrile-
Water 17：83 be mobile phase；Detection wavelength is 283nm；Number of theoretical plate is calculated by aurantiamarin peak should be not less than 5000；
The preparation of reference substance solution takes aurantiamarin reference substance appropriate, accurately weighed, adds methanol that every 1ml is made molten containing 15 μ g
Liquid, to obtain the final product；
The preparation of test solution takes this product, finely ground, crosses No. five sieves, takes 2.0g, accurately weighed, puts in conical flask with cover,
Precision adds 10% methanol 50ml, and weighed weight, is ultrasonically treated 45 minutes, takes out, lets cool, then weighed weight, with 10% methanol
Supply the weight of less loss, shake up, filter, take subsequent filtrate, to obtain the final product；
Determination method difference is accurate to draw reference substance solution and each 10 μ l of test solution, injects liquid chromatograph, measures, i.e.,
.
Every bag of this product is containing avicenna pricklyash root with aurantiamarin (C28H34O15) meter, 2.0mg must not be less than.
Methodology validation：
2 optimal extraction technologies
2.1 different extracting modes are investigated
Guanule contg. ' Jigucao particle 2.0g is taken, two parts, precision weighs, and precision adds methanol 25ml, weighed weight, a ultrasound
30min is extracted, another is heated to reflux 30min, lets cool, and mends weight, takes subsequent filtrate, crosses 0.45 μm of miillpore filter, sample introduction, the result is shown in
Table 1.The result shows that heating and refluxing extraction is more slightly higher than the content of ultrasonic extraction, through SPSS single factor analysis, conspicuousness is displayed without
Difference, it is contemplated that ultrasonic extraction facilitates the advantages that easy to operate, environmentally friendly, selects ultrasonic extraction.
1 Different Extraction Method comparision contents of table
2.2 different extraction solvents are investigated
Guanule contg. ' Jigucao particle 2.0g is taken, two parts, precision weighs, a accurate addition absolute ethyl alcohol 25ml, weighed weight,
Another precision adds methanol 25ml, and ultrasonic extraction 30min, lets cool, and mends weight, takes subsequent filtrate, crosses 0.45 μm of miillpore filter, into
Sample.The result shows that the extraction effect of methanol is better than absolute ethyl alcohol.50% methanol, the extraction effect of 10% methanol are further investigated,
The result shows that the content highest of 10% methanol extraction, the results are shown in Table 2.
Influence of the different extraction solvents of table 2 to content
2.3 orthogonal test
According to single factor exploration as a result, selection methanol concentration, solvent is measured again, extraction time is investigation factor, set 3 respectively
Level, according to L (34) (it is blank assay to have a level) orthogonal arrage tested, 3 are shown in Table, orthogonal experiments are shown in Table 4 Hes
Table 5.
3 orthogonal test factor level table of table
4 orthogonal experiments of table
5 analysis of variance table of table
As seen from the above table, it is methanol concentration (A) that each factor influences size order to assay result>Solvent dosage (B)>
Extraction time (C), above three factor there are no significant difference.Optimal experimental program is A1B3C2 obtained by orthogonal test, is considered
There is the situation of super balance range in the solvent that 100ml is added into conical flask.And the average difference of two factors of 50ml and 100ml
4.566, variance analysis shows do not have significant difference in solvent dosage group.Therefore, the optimal testing program of comprehensive income is：
The 10% methanol ultrasonic extraction 45min of A1B2C2, i.e. 50ml.
3 system suitabilities
3.1 specificities are tested
Lack avicenna pricklyash root feminine gender solution, Guanule contg. ' Jigucao particle test solution, aurantiamarin reference substance solution difference sample introduction point
Analysis, is shown in Fig. 4~6, as a result aurantiamarin peak with it is similar it is other (separating degree is more than 1.5) is kept completely separate into swarming, and lack avicenna pricklyash root
Negative sample solution is noiseless at the identical appearance time of aurantiamarin reference substance.
3.2 linear relationships are investigated
Take aurantiamarin reference substance solution and take 1ml aurantiamarin reference substance solutions, be respectively placed in 2ml, 5ml, 10ml, 25ml,
In the volumetric flask of 50ml, 100ml, methanol is added to shake up, be measured by above-mentioned chromatographic condition, confronted with peak area (Y) to scale
Measure concentration (X) and carry out linear regression.The calibration curve equation of aurantiamarin is Y=21838.6976X+3169.8100.01 (R2=
1.0000) it is that linear relationship is good in the range of 1.2904~129.0362 μ g/ml in concentration, as shown in Figure 3.
3.3 precision test
Same reference substance solution (aurantiamarin concentration is 12.9036 μ g/ml) is taken, precision draws 10 μ l injection high-efficient liquid phase colors
Spectrometer, measures in accordance with the law, the results are shown in Table 6, peak area RSD is 1.11%, it is seen that instrument precision is good.
6 Precision test result of table
3.4 repetitive test
6 parts of same batch sample (QL002) is taken, it is accurately weighed, test solution is prepared, peak area is recorded and calculating contains
Amount, is shown in Table 7, and aurantiamarin average content is 132.2 μ g/g, RSD 0.53%, shows that this method repeatability is good.
7 repetitive test result (n=6) of table
3.5 sample-adding recovery tests
Sample (lot number QL002) 1.0g of known content is taken, it is accurately weighed, 6 parts are taken altogether, is put in conical flask with cover, respectively
Precision adds reference substance solution (7.9186 μ g/ml) 50ml, by method time-and-motion study under " 3.3 " item, calculates the rate of recovery, the result is shown in
Table 8.
8 sample recovery rate result of the test (n=6) of table
3.6 serviceability test
3.6.1 stability test
Same test solution (QL002) 2.0g is taken, confession is prepared according to the preparation method of test solution under assay item
Test sample solution, is measured in different time respectively, the results are shown in Table 9, test solution is stablized in 5h.
9 stability test result of table
3.6.2 column temperature is investigated
Take with two parts of test solution (QL002) 2.0g, prepared according to the preparation method of test solution under assay item
Test solution, is respectively measured different column temperatures, the results are shown in Table 10.Result of the test shows in 25~40 DEG C to content
Measurement result does not have a significant impact, and is preferably 30 DEG C.
Influence of the different column temperatures of table 10 to assay
3.6.3 flow velocity is investigated
Same test solution (QL002) 2.0g is taken, confession is prepared according to the preparation method of test solution under assay item
Test sample solution, is respectively measured tri- flow velocitys of 0.8ml/min, 1.0ml/min, 1.2ml/min, the results are shown in Table 11.Experiment
It is preferably 1.0ml/min the result shows that not having a significant impact to assay result in 0.8~1.2ml/min flow velocitys.
The influence different in flow rate to assay result of table 11
3.6.4 different wave length is investigated
Same test solution (QL002) 2.0g is taken, confession is prepared according to the preparation method of test solution under assay item
Test sample solution, is measured in five wavelength of 278nm, 281nm, 283nm, 285nm, 287nm, the results are shown in Table 12 respectively.Experiment
It is preferably 283nm the result shows that not having a significant impact to assay result in 283 ± 5nm.
Influence of 12 different wave length of table to assay result
3.6.5 mobile phase proportion of composing investigates experiment
Methanol-water flow phase system is investigated, the results show is compared with acetonitrile-water flow phase system, testing compound peak
Shape is wide, and theoretical cam curve is lower slightly, therefore selects acetonitrile-water flow phase system.
When mobile phase ratio is acetonitrile-water (18:82), adjacent peak causes to do to peak to be measured in the results show test solution
Disturb, separating degree is less than 1.5, does not reach assay requirement.When mobile phase ratio is acetonitrile-water (16:84) when, with adjacent peak point
From good, aurantiamarin reference substance appearance time is delayed, and greatly increases analysis time, and efficiency reduces.Ensureing aurantiamarin reference substance
On the premise of meeting liquid phase systems adaptability, and there is the suitable analysis time, therefore selective flow is mutually acetonitrile-water (17:83).
The assay of 3.7 10 batches of Zishenningshenwan test samples
10 batches of test samples of Guanule contg. ' Jigucao particle are taken, are prepared according to the preparation method of test solution under assay item for examination
Product solution, records peak area and calculates content, the results are shown in Table shown in 13.
13 10 batches of Guanule contg. ' Jigucao particle content measuring results of table
Claims (2)
1. a kind of quality determining method of Guanule contg. ' Jigucao particle, it is characterised in that including thin-layer chromatography Qualitive test and orange peel
The content detection of glycosides：
（1）The thin-layer chromatography Qualitive test of hypericum japonicum：
This product 3 g is taken, it is finely ground, add 25 ml of water, 20-40 minutes ultrasonic, filtration, adds dilute hydrochloric acid tune pH 1 ~ 2, extracted with ethyl acetate
Take 1-3 times, every time 20 ml, combined ethyl acetate extract, is evaporated, and residue adds ethanol 1ml to make dissolving, as test solution；
1 g of hypericum japonicum control medicinal material separately is taken, adds 25 ml of water, is heated to reflux 20-40 minutes, is filtered, from " adding dilute hydrochloric acid tune pH 1 ~ 2 "
Hypericum japonicum control medicinal material solution is made in the same way of, it is spare；Tested according to thin-layered chromatography, draw each 5 ~ 10 μ l of above two solution, point
Other point is on same silica gel g thin-layer plate, with volume ratio dichloromethane：Ethyl acetate：Methanol：Formic acid：Water is 12:10:2:1:1 is
Solvent, is unfolded, and takes out, dries, spray with alchlor test solution, heats 5 ~ 10min at 105 DEG C, puts and examined under 365nm ultraviolet lamps
Depending on；In test sample chromatography, on position corresponding with control medicinal material chromatography, same color spot；
（2）The assay of aurantiamarin：
It is measured using high performance liquid chromatography, specific method and step are as follows：
Chromatographic condition is filler with octadecylsilane chemically bonded silica with system suitability；With volume ratio acetonitrile-water 17：
83 be mobile phase；Detection wavelength is 283nm, flow velocity 1.0ml/min；Number of theoretical plate is calculated by aurantiamarin peak to be not less than
5000；
The preparation of reference substance solution takes aurantiamarin reference substance appropriate, accurately weighed, adds methanol that the solution that every 1ml contains 15 μ g is made,
To obtain the final product；
The preparation of test solution takes this product, finely ground, crosses No. five sieves, takes 2.0g, accurately weighed, puts in conical flask with cover, accurate
10% methanol 50ml is added, weighed weight, is ultrasonically treated 45 minutes, takes out, lets cool, then weighed weight, is supplied and subtracted with 10% methanol
The weight of mistake, shakes up, and filtration, takes subsequent filtrate, to obtain the final product；
Determination method difference is accurate to draw reference substance solution and each 10 μ l of test solution, injects liquid chromatograph, measures, to obtain the final product.
2. the quality determining method of a kind of Guanule contg. ' Jigucao particle according to claim 1, it is characterised in that described（3）
In, test solution is prepared as：This product is taken, it is finely ground, No. five sieves are crossed, take 2.0g, it is accurately weighed, put in conical flask with cover, essence
10% methanol 50ml of close addition, weighed weight, is ultrasonically treated 45 minutes, takes out, lets cool, then weighed weight, is supplied with 10% methanol
The weight of less loss, shakes up, and filtration, takes subsequent filtrate, to obtain the final product.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111024875A (en) * | 2019-12-31 | 2020-04-17 | 广东药科大学 | Construction method of abrus cantoniensis hance amide component liquid chromatography fingerprint |
| CN114814068A (en) * | 2022-05-13 | 2022-07-29 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus cantoniensis hance and Abrus mollis |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1529163A (en) * | 2003-10-16 | 2004-09-15 | 中山大学 | Method for constituting TLC finger-print pyrogram of hypericum-japonicum medicinal herbs and its standard finger-print pyrogram |
| WO2012167457A1 (en) * | 2011-06-07 | 2012-12-13 | Su Weiwei | Use of hypericum japonicum thunb. general flavone in preparing medicament for treating hepatic fibrosis |
| CN103585409A (en) * | 2013-11-18 | 2014-02-19 | 镇江天和生物技术有限公司 | Polygonum cuspidatum and glossy privet fruit mixture and preparation method thereof |
-
2017
- 2017-12-05 CN CN201711269360.1A patent/CN108037234B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1529163A (en) * | 2003-10-16 | 2004-09-15 | 中山大学 | Method for constituting TLC finger-print pyrogram of hypericum-japonicum medicinal herbs and its standard finger-print pyrogram |
| WO2012167457A1 (en) * | 2011-06-07 | 2012-12-13 | Su Weiwei | Use of hypericum japonicum thunb. general flavone in preparing medicament for treating hepatic fibrosis |
| CN103585409A (en) * | 2013-11-18 | 2014-02-19 | 镇江天和生物技术有限公司 | Polygonum cuspidatum and glossy privet fruit mixture and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| QING MU 等: "Antibacterial Effects of Hypericum ascyron. and Hypericum japonicum. Against Multidrug-Resistant Staphylococcus aureus.", 《PHARMACEUTICAL BIOLOGY》 * |
| 侯小涛 等: "高效液相色谱法同时测定鸡骨草肝炎颗粒中原儿茶酸、原儿茶醛及没食子酸的含量", 《中国医院药学杂志》 * |
| 程静 等: "鸡骨草肝炎颗粒的薄层鉴别和绿原酸、槲皮素的测定", 《华西药学杂志》 * |
| 莫少红 等: "HPLC法测定鹰不泊药材中橙皮苷的含量", 《中药新药与临床药理》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111024875A (en) * | 2019-12-31 | 2020-04-17 | 广东药科大学 | Construction method of abrus cantoniensis hance amide component liquid chromatography fingerprint |
| CN111024875B (en) * | 2019-12-31 | 2022-06-17 | 广东药科大学 | Construction method of abrus cantoniensis hance amide component liquid chromatography fingerprint |
| CN114814068A (en) * | 2022-05-13 | 2022-07-29 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus cantoniensis hance and Abrus mollis |
| CN114814068B (en) * | 2022-05-13 | 2023-11-17 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus herb and abrus herb |
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