CN104777249B - The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf - Google Patents
The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf Download PDFInfo
- Publication number
- CN104777249B CN104777249B CN201510160159.4A CN201510160159A CN104777249B CN 104777249 B CN104777249 B CN 104777249B CN 201510160159 A CN201510160159 A CN 201510160159A CN 104777249 B CN104777249 B CN 104777249B
- Authority
- CN
- China
- Prior art keywords
- amygdaloside
- solution
- cough syrup
- reference substance
- content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention discloses a kind of method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf, it includes the preparation process of reference substance solution, the preparation process of need testing solution and high-performance liquid chromatogram determination step, determine that the every 1ml of product must not be less than 1.2mg containing amygdaloside, the present invention is measured to the amygdaloside content controlling cough syrup of loquat leaf product quality using high performance liquid chromatography, detection compared to no assay item in existing standard, the inherent quality of medicine directly can be controlled by the content detecting amygdaloside, and in this assay method, amygdaloside good separating effect, stability and accuracy are high, reproducible, the quality of cough syrup of loquat leaf product so can be better controled over.
Description
Technical field
The present invention relates to a kind of detection method of medicine is and in particular to measure effective ingredient Semen Armeniacae Amarum in cough syrup of loquat leaf
The method of glycosides content.
Background technology
Cough syrup of loquat leaf is conventional Chinese medicine preparation, has lung heat clearing, cough-relieving, effect of resolving sputum, for cough ant phlegm, bronchus
Scorching treatment, due to the existing quality standard of cough syrup of loquat leaf (《The Sanitation Ministry medicine standard》Traditional Chinese medicine historical preparation the 9th)
(WS3-B-1702-94) only define routine examination, specially：(1) take this product cough syrup of loquat leaf 1mL, add water 10mL, strongly
Shaking, has persistent foam to generate；(2) take this product 10mL, hydro-oxidation sodium test solution makes into alkalescence, extracted with chloroform 10mL, point
Take chloroform layer, use 1% sulphuric acid solution extraction, acid solution adds bismuth potassium iodide test solution, generate brown precipitate.This standard no assay item
Detection, it is impossible to efficiently control the inherent quality of medicine, according to related request, needs to revise and enlarge content measuring standard to said preparation,
Control product quality with more preferable.
The prescription of cough syrup of loquat leaf consists of：Folium Eriobotryae 391g, Rhizoma Cynanchi Stauntonii 125g, Cortex Mori 234g, Radix Platycodoniss 78g, the Radix Stemonae
94g, Oleum menthae 3mL, wherein Folium Eriobotryae are party's principal agent, are confirmed as content detection object.
The assay composition with ursolic acid and oleanolic acid as Folium Eriobotryae for the Chinese Pharmacopoeia 2010 editions, yet with preparation work
The content of the loss that skill is led to, ursolic acid and oleanolic acid is atomic, and therefore ursolic acid and oleanolic acid are not suitable as this system
The content's index of agent.Document shows that Folium Eriobotryae contains amygdaloside, ursolic acid, oleanolic acid, tartaric acid, citric acid, malic acid, tan
The compositions such as matter, vitamin B and vitamin C, and amygdaloside has relieving cough and resolving phlegm effect, is the active component of Folium Eriobotryae, is suitable for
As assay object.
Therefore, in the urgent need to researching and developing a kind of method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf, come
Standard as evaluation cough syrup of loquat leaf quality has great importance.
Content of the invention
For overcoming the shortcomings of above-mentioned existing background technology, it is an object of the invention to provide one kind can better control over cough-relieving
The assay method of the amygdaloside content of loquat syrup for treating product quality.
The purpose of the present invention is reached by following measure：Effective ingredient hardship Fructus Pruni in a kind of mensure cough syrup of loquat leaf
The method of core glycosides content, comprises the following steps：
(1) preparation of reference substance solution：Take amygdaloside reference substance appropriate, accurately weighed, plus mobile phase makes every 1mL and contains
The solution of 80 μ g, obtains final product reference substance amygdaloside solution；
(2) preparation of need testing solution：Measure this product cough syrup 5mL, put 10mL volumetric flask, plus ethanol is settled to capacity
Bottle scale, shakes up, with filter paper filtering, takes subsequent filtrate 1mL, put 10mL volumetric flask, be diluted to volumetric flask scale with mobile phase, shake
Even, with 0.45 μm of membrane filtration, take subsequent filtrate, subsequent filtrate is need testing solution；
(3) use high-performance liquid chromatogram determination：With octadecylsilane chemically bonded silica as filler, wherein flow rate of mobile phase is
1.0mL/min, column temperature is 30 DEG C, and Detection wavelength is 210nm, and number of theoretical plate is not less than 4000 based on amygdaloside peak, inhales respectively
Take reference substance solution and each 20 μ l of need testing solution, inject chromatograph of liquid, measure, obtain the every 1mL of need testing solution and contain Semen Armeniacae Amarum
Glycosides must not be qualified less than 1.2mg；
In described mobile phase, the volume ratio of each composition is：Methanol：Water=20: 80.
When further, using high-performance liquid chromatogram determination, pillar height 250mm of chromatographic column used, internal diameter 4.6mm, filling
Agent particle diameter is 5 μm.
The beneficial effects of the present invention is：The present invention uses high performance liquid chromatography to control cough syrup of loquat leaf product matter
The amygdaloside content of amount is measured, and compared to the detection of no assay item in existing standard, the present invention can directly pass through
The inherent quality to control medicine for the content of detection amygdaloside, and in this assay method, amygdaloside good separating effect, surely
Qualitative and accuracy is high, reproducible, so can better control over the quality of cough syrup of loquat leaf product.
Brief description
Fig. 1 is amygdaloside reference substance chromatogram；
Fig. 2 is test sample chromatogram；
Fig. 3 is negative control chromatogram；
Fig. 4 is amygdaloside standard curve.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples, but they are not constituted to the present invention
Restriction, only for example.Simultaneously by illustrate advantages of the present invention will become clearer from easy to understand.
Embodiment
The present invention provides a kind of method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf, walks including following
Suddenly：
(1) preparation of reference substance solution：Take amygdaloside reference substance appropriate, accurately weighed, plus mobile phase makes every 1mL and contains
The solution of 80 μ g, obtains final product reference substance amygdaloside solution；
(2) preparation of need testing solution：Measure this product cough syrup 5mL, put 10mL volumetric flask, plus ethanol is settled to capacity
Bottle scale, shakes up, with filter paper filtering, takes subsequent filtrate 1mL, put 10mL volumetric flask, be diluted to volumetric flask scale with mobile phase, shake
Even, with 0.45 μm of membrane filtration, take subsequent filtrate, subsequent filtrate is need testing solution；
(3) use high-performance liquid chromatogram determination：With octadecylsilane chemically bonded silica as filler, wherein flow rate of mobile phase is
1.0mL/min, column temperature is 30 DEG C, and Detection wavelength is 210nm, and number of theoretical plate is not less than 4000 based on amygdaloside peak, inhales respectively
Take reference substance solution and each 20 μ l of need testing solution, inject chromatograph of liquid, measure, obtain the every 1mL of need testing solution and contain Semen Armeniacae Amarum
Glycosides must not be qualified less than 1.2mg；
In described mobile phase, the volume ratio of each composition is：Methanol：Water=20：80；Pillar height 250mm of chromatographic column used, interior
Footpath 4.6mm, filler particle diameter is 5 μm.
Correlation test is reported as follows：
1st, instrument and reagent
DIONEX high performance liquid chromatograph, Μ VD170 μ LC UV-detector, (Beijing Sai Duolisi balance has electronic balance
Limit company), Welch (moon rising sun) Μ ltimate XB-C18 post (250mm × 4.6mm, 5 μm).Cough syrup of loquat leaf (rectify by Hubei
Pharmaceutcal corporation, Ltd produces, lot number 130101,130102,130103)；Amygdaloside reference substance (grind by Chinese food drug assay
Study carefully institute, lot number 110820-201004, for assay)；HPLC water is distilled water (laboratory self-control), and methanol is chromatograph
Pure, it is pure that ethanol (Chemical Reagent Co., Ltd., Sinopharm Group), methanol (Chemical Reagent Co., Ltd., Sinopharm Group) etc. are analysis.
The preparation of 1.1 reference substance solution
Take amygdaloside reference substance appropriate, accurately weighed, plus mobile phase makes the solution that every 1mL contains 80 μ g, obtains final product comparison
Product amygdaloside solution；
The preparation of 1.2 reference substance stock solutions
Precision weighs amygdaloside reference substance 12.82mg to 25mL volumetric flask, uses mobile phase constant volume, obtains reference substance and stock
Liquid.
The preparation of 1.3 need testing solutions
Precision measures this product 5mL, puts 10mL volumetric flask, plus ethanol, to scale, shakes up, filtration, takes subsequent filtrate 1mL, with stream
Dynamic phase dilution, to scale, shakes up, 0.45 μm of filter membrane filtration, takes subsequent filtrate, obtains final product.
The preparation of 1.4 negative control solutions
Existing cough syrup of loquat leaf preparation method (《The Sanitation Ministry medicine standard》Traditional Chinese medicine historical preparation the 9th)) as follows：
Folium Eriobotryae 391g, Rhizoma Cynanchi Stauntonii 125g, Cortex Mori 234g, Radix Platycodoniss 78g, Radix Stemonae 94g, Oleum menthae 3ml, above Six-element, removes
Outside Oleum menthae, Folium Eriobotryae, Radix Platycodoniss, the Radix Stemonae, Cortex Mori are ground into coarse powder.Try with the percolation under extractum item according to fluid extract
Test, be that solvent carries out percolation with 50～60% ethanol, collect liquid of filtering, reclaim ethanol, be condensed into 1：2 thick paste；Rhizoma Cynanchi Stauntonii adds water pan-fried
Boil 2 times, 1 hour every time, collecting decoction, filtration, it is condensed into 1：2 thick paste, places 12 hours, filtration, after the appropriate dilution that adds water,
With sucrose 900g, benzoic acid 3.75g, citric acid 0.75g, boil and make dissolving, filter while hot, add above-mentioned alcohol medicinal extract, stir evenly, plus
Enter Oleum menthae and essence 4.05ml (being dissolved in ethanol in proper amount), add water to 1500ml, stir evenly, fill, obtain final product.
Make the negative controls without Folium Eriobotryae by above-mentioned prescription and technique, it is negative right to make by the method under 1.3
According to solution.
2. method and result
2.1 chromatographic system compatibility tests
Μ ltimate XB-C18 post (250mm × 4.6mm, 5 μm)；With methanol-water (volume ratio 20: 80) as mobile phase；
Flow velocity：1.0mL/min；Column temperature：30℃；Detection wavelength：210nm；Take respectively the reference substance solution preparing, need testing solution,
Negative control solution sample introduction, sample size is all 20 μ L, and result such as Fig. 1 to 3, Fig. 1 amygdaloside retention time is in 16.715min；
Corresponding with Fig. 1 amygdaloside reference substance retention time, the amygdaloside chromatographic peak in Fig. 2 is kept completely separate with other impurities peak,
Good separating effect, the selection of therefore the adopted chromatographic column of explanation experiment, mobile phase and proportioning, retention time is appropriate, chromatostrip
Part is good；In Fig. 3, negative control solution does not show amygdaloside at retention time identical with reference substance solution and need testing solution
Chromatographic peak, therefore think noiseless to measuring.
2.2 standard curve
Take amygdaloside reference substance storing solution, series concentration, the accurate reference substance drawing above-mentioned variable concentrations are made in dilution
The each 20 μ L of solution, inject high performance liquid chromatograph, measure chromatographic peak area according to 2.1 conditions, and related concentrations and peak area are shown in Table 1,
With the concentration of reference substance solution as abscissa, peak area is vertical coordinate, draws standard curve, and regression equation is：Y=
0.2716x 1.36466, r=0.9981, is shown in Fig. 4, and result shows in amygdaloside in 9.61～153.8 μ g/mL linear relationships
Well.
Table 1 linear relationship
2.3 precision test
Accurate absorption reference substance solution 20 μ L, is measured by 2.1 chromatographic conditions, repeats sample introduction 5 times, measure and absorb peak area,
The results are shown in Table 2, RSD is 1.92%, shows that instrument precision is good, the numerical value that the method measures is accurate, credible.
Table 2 Precision test result
2.4 replica test
Take lot number to be 5 parts of 120102 Folium Eriobotryae cough syrup, measure by need testing solution preparation method and 2.1 chromatographic conditions,
Measure it and absorb peak area, the results are shown in Table 3, RSD is 2.62%, show that the repeatability of the method is good.
Table 3 replica test result
2.5 stability test
Take with portion need testing solution, respectively at 0h, 1h, 4h, 8h, 12h, 24h sample introduction 6 times respectively, measure its absworption peak
Area, result RSD is 1.19%, shows that solution is stable in 24h, the results are shown in Table 4.
Table 4 stability test result
2.6 average recovery
Accurate 5 parts of same a collection of Folium Eriobotryae cough syrup (lot number is 131002) drawing known content, every part of syrup 1mL, respectively
It is separately added into amygdaloside 1795.5 μ g in part, is settled to 2mL with ethanol, then press need testing solution preparation method and filter, take continuous
Filtrate 1mL flowing phase dilution is simultaneously settled to 10mL, 0.45 μm of filter membrane filtration, takes subsequent filtrate to prepare test liquid, by 2.1 chromatostrips
Part measures, calculates, and the results are shown in Table 5, and amygdaloside average recovery rate is 95.19%.
Table 5 is loaded recovery test result
3. assay and limit are worked out
Prepared by need testing solution to 3 batch samples, measures by 2.1 chromatographic conditions, calculate content, the results are shown in Table 6,3 batch samples
The content of every 100mL amygdaloside is 158.39mg to 161.57mg.
Table 6 Folium Eriobotryae cough syrup sample size measurement result
Above-mentioned three batch samples detect for sample, detect for full-page proof in actual production, and the content according to 3 batch samples combines and produces
Reality, determines this product every 1mL (C containing amygdaloside20H27NO11) amount must not be less than 1.20mg, determine in cough syrup of loquat leaf bitter
Semen Armeniacae Amarum glycosides content is more than or equal to 1.20mg/mL, illustrates that it is the certified products by existing cough syrup of loquat leaf preparation method gained；If bitter
Semen Armeniacae Amarum glycosides content is less than 1.20mg/mL, illustrates that this cough syrup of loquat leaf is unqualified.
Conclusion：Each item data that above method is investigated all meets the pertinent regulations of quality standards in Chinese drugs research, text institute
Method of stating, be applied to the content measuring amygdaloside cough syrup of loquat leaf, therefore can directly be passed through to detect the content of amygdaloside
To control the inherent quality of medicine, and in this assay method, amygdaloside good separating effect, stability and accuracy height, weight
Renaturation is good, shows party's forensic science, reasonable, feasible.
Other unspecified parts belong to prior art.
Claims (1)
1. measure cough syrup of loquat leaf in effective ingredient amygdaloside content method it is characterised in that：Comprise the following steps：
Step one：The preparation of reference substance solution：Take amygdaloside reference substance appropriate, accurately weighed, plus mobile phase makes every 1mL and contains
The solution of 80 μ g, obtains final product reference substance amygdaloside solution；
Step 2：The preparation of need testing solution：Measure this product cough syrup 5mL, put 10mL volumetric flask, plus ethanol is settled to capacity
Bottle scale, shakes up, with filter paper filtering, takes subsequent filtrate 1mL, put 10mL volumetric flask, be diluted to volumetric flask scale with mobile phase, shake
Even, with 0.45 μm of membrane filtration, take subsequent filtrate, subsequent filtrate is need testing solution；
Step 3：Use high-performance liquid chromatogram determination：With octadecylsilane chemically bonded silica as filler, wherein flow rate of mobile phase is
1.0mL/min, column temperature is 30 DEG C, and Detection wavelength is 210nm, and number of theoretical plate is not less than 4000 based on amygdaloside peak, inhales respectively
Take reference substance solution and each 20 μ l of need testing solution, inject chromatograph of liquid, measure, obtain final product the amygdaloside in need testing solution
Content；
In described mobile phase, the volume ratio of each composition is：Methanol：Water=20：80；
In described step 3, during using high-performance liquid chromatogram determination, pillar height 250mm of chromatographic column used, internal diameter 4.6mm, filling
Agent particle diameter is 5 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510160159.4A CN104777249B (en) | 2015-04-07 | 2015-04-07 | The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510160159.4A CN104777249B (en) | 2015-04-07 | 2015-04-07 | The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104777249A CN104777249A (en) | 2015-07-15 |
| CN104777249B true CN104777249B (en) | 2017-03-01 |
Family
ID=53618849
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510160159.4A Expired - Fee Related CN104777249B (en) | 2015-04-07 | 2015-04-07 | The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104777249B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891381A (en) * | 2016-04-22 | 2016-08-24 | 广西壮族自治区梧州食品药品检验所 | Method using ASE to extract amygdalin in semen persicae |
| CN105929059A (en) * | 2016-04-22 | 2016-09-07 | 广西壮族自治区梧州食品药品检验所 | Determination method for amygdalin in peach seed |
| CN106290649A (en) * | 2016-08-31 | 2017-01-04 | 天津中新药业研究中心 | A kind of method of quality control of removing heat from the lung and dissipating phlegm antitussive medicine |
| CN108267515B (en) * | 2016-12-30 | 2020-10-02 | 广州中医药大学第一附属医院 | Method for detecting content of D-amygdalin and/or L-amygdalin in substance to be detected |
| CN106841467A (en) * | 2017-03-03 | 2017-06-13 | 四川德成动物保健品有限公司 | The high-efficiency liquid chromatography method for detecting of amarogentin in a kind of female biochemistry mixture of benefit |
| CN111175392B (en) * | 2019-12-31 | 2022-05-03 | 江西杏林白马药业股份有限公司 | Quality control method of cough-relieving loquat syrup |
| CN112345676B (en) * | 2020-12-07 | 2022-06-07 | 葵花药业集团湖北武当有限公司 | HPLC method for simultaneously detecting 6 active ingredients in fritillaria cirrhosa lung-heat-clearing syrup |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102297926B (en) * | 2011-07-12 | 2013-08-28 | 刘金伶 | Rapid TLC identification and three-component simultaneous quantitative determination method for antitussive tablets |
| CN103604885A (en) * | 2013-11-21 | 2014-02-26 | 天津中新药业集团股份有限公司第六中药厂 | Method for determining quality of cough-relieving fritillary and loquat leaf drop pills |
-
2015
- 2015-04-07 CN CN201510160159.4A patent/CN104777249B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104777249A (en) | 2015-07-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104777249B (en) | The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf | |
| CN105606734A (en) | Method for detecting honeysuckle flower and lonicerae flos medicinal materials through rapid resolution liquid chromatography | |
| CN105259295B (en) | Quality detection method for ginseng, cassia twig and poria cocos oral solution | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN104897787B (en) | A kind of method of six kinds of active component in NIUHUANG NINGGONG PIAN of mensuration simultaneously | |
| CN106324161B (en) | Treat the quality determining method of the Chinese medicine composition of diabetic nephropathy | |
| CN106370738B (en) | Fingerprint quality determination method for periplaneta americana medicinal material | |
| CN105067747B (en) | The detection method of Styrax flavour of a drug ingredients fingerprint and its application in a kind of Heart pill of Musk | |
| CN104764820A (en) | Method for determining content of active ingredients such as ephedrine hydrochloride and pseudoephedrine hydrochloride in pinellia ternata syrup | |
| CN101703611A (en) | Quality detection method of Chinese angelica oral liquid for benefiting blood | |
| CN103926332B (en) | A kind of Ultra Performance Liquid Chromatography method of uridine, guanosine and adenosine content in Simultaneously test pinellia tuber extract | |
| CN112666277B (en) | HPLC (high Performance liquid chromatography) characteristic spectrum construction and detection method for rhizoma cyperi medicinal materials, decoction pieces, standard decoction and formula granules | |
| CN102998383B (en) | Method for testing content of main components in Rhodiola rosea extracts | |
| CN101543545B (en) | Traditional Chinese medicine preparation for curing rhinitis and quality control method thereof | |
| CN108709949B (en) | Detection method of fermented cordyceps sinensis powder Cs-4 traditional Chinese medicine decoction pieces | |
| CN102552515A (en) | Quality detection method for blood-nourishing Chinese angelica syrup | |
| CN104897811B (en) | A kind of detection method of Moschus 'Xintongning ' | |
| CN101703610A (en) | Quality detection method of Qingnao antihypertensive tablet | |
| CN106680414A (en) | Detection method of compound ardisia japonica tablet | |
| CN103713067B (en) | Ultra-high performance liquid chromatography method for determining content of rheum lhasaense | |
| CN101987115A (en) | Jade screen oral preparation as well as preparation method and detection method thereof | |
| CN105486761A (en) | Method for determining scutelloside content in traditional Chinese medicine granules | |
| CN108490095A (en) | A kind of method of Multiple components assay in pilose gerbera herb medicinal material | |
| CN110231418B (en) | Method for separating and measuring contents of various components in Huoxiang Zhengqi oral liquid by HPLC method | |
| CN104597168B (en) | Ramulus Mori refines decoction pieces content assaying method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170301 Termination date: 20200407 |