CN113916999A - HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule - Google Patents
HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule Download PDFInfo
- Publication number
- CN113916999A CN113916999A CN202110445464.3A CN202110445464A CN113916999A CN 113916999 A CN113916999 A CN 113916999A CN 202110445464 A CN202110445464 A CN 202110445464A CN 113916999 A CN113916999 A CN 113916999A
- Authority
- CN
- China
- Prior art keywords
- solution
- tortoise
- detection method
- kidney tonifying
- hesperidin
- 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.)
- Withdrawn
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
Abstract
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an HPLC (high performance liquid chromatography) detection method for a dried orange peel component in a tortoise-deer kidney tonifying capsule, which comprises the steps of (1) chromatographic condition and system applicability test (2) solution preparation (3) determination and the like. The HPLC detection method for the dried orange peel component in the tortoise and deer kidney tonifying capsule has the advantages of simple operation, high detection efficiency, small error, good reproducibility, strong specificity, stable detection method, fewer influenced factors and wide applicability; the specificity, stability, precision, accuracy and the like of the method all meet the requirement of methodology verification, and the method has strong practicability.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an HPLC (high performance liquid chromatography) detection method for a dried orange peel component in a tortoise-deer kidney tonifying capsule.
Background
The Guilu kidney-tonifying capsule is a Chinese patent medicine prescription preparation, has the effects of strengthening the muscles and bones, replenishing qi and blood, tonifying the kidney and strengthening yang, and is collected in the seventeenth national food and drug administration standard YBZ00672012 of the Chinese patent medicine prescription preparation issued by the ministry of health. The traditional Chinese medicine composition is mainly used for treating weakness, mental fatigue, soreness and weakness of waist and legs, dizziness, kidney deficiency, cold sperm, hyposexuality, nocturia, amnesia and insomnia in clinic. Because of its definite curative effect, it is widely used in clinic, and its prescription composition is made up by using dodder seed, epimedium, dipsacus root, cynomorium songaricum, cibotium root, spiny jujube kernel, prepared flowery knotweed root, honey-fried licorice root, tangerine peel, deerhorn glue, cooked rehmannia root, tortoise-shell glue, cherokee rose fruit, astragalus root, Chinese yam and raspberry fruit.
The dried orange peel is one of the main effective components of the formula of the tortoise plastron and deer kidney tonifying capsule, and has the effects of regulating qi, strengthening spleen, eliminating dampness and phlegm. It is mainly used for treating abdominal distention, anorexia, vomiting, diarrhea, cough and excessive phlegm. The main effective components of the traditional Chinese medicine composition comprise volatile oil and flavonoid substances (mainly hesperidin), and the content of the hesperidin is generally determined to be used as a quality standard for evaluating the dried orange peel. In the existing standard of Guilu Bushen capsule, the drug standard promulgated by the Ministry of health, traditional Chinese medicine finished formula preparation, the seventeenth national food and drug administration Standard YBZ00672012, the content of pericarpium Citri Tangerinae component hesperidin is not controlled, and only icariin is used as a quality control index to control the content of herba Epimedii, which is not comprehensive enough and unreasonable.
The invention aims to provide an HPLC detection method for a dried orange peel component in a tortoise and deer kidney tonifying capsule aiming at the defects in the prior art, and the method for detecting the flavonoid hesperidin in the dried orange peel has the advantages of simple operation, high detection efficiency, small error, good reproducibility, strong specificity, more stable detection method and wide applicability, and can effectively control the quality of the tortoise and deer kidney tonifying capsule.
Disclosure of Invention
In order to achieve the purpose, the invention adopts the following technical scheme:
an HPLC detection method for a dried orange peel component in a tortoise plastron and deer kidney tonifying capsule comprises the following steps:
(1) chromatographic conditions and system applicability test:
octadecylsilane chemically bonded silica is used as a filler, and a chromatographic column comprises the following components: waters
T3C 18 column, 4.6X 250mm, 5 μm or equivalent column, mobile phase: acetonitrile-water (22: 78). Flow rate: 1.0ml/min, detection wavelength: 283nm, column temperature: 35 ℃, sample introduction: 10 mul, the theoretical plate number is not lower than 5000 calculated according to the hesperidin peak.
(2) Solution preparation:
preparation of a test solution: taking the contents of the product with different filling amounts, mixing uniformly, grinding, taking about 1g, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of methanol, sealing the plug, weighing, heating and refluxing for 3 hours, cooling, weighing again, supplementing the lost weight with methanol, shaking up, filtering, precisely weighing 20ml of subsequent filtrate, recovering the solvent to dryness, adding a proper amount of methanol into the residue for dissolving, transferring to a 5ml measuring flask, adding methanol to scale, shaking up, filtering, taking the subsequent filtrate, centrifuging at 5000 r/min for 5min, and filtering with a 0.45 mu m filter membrane to obtain the sample solution.
Preparing a reference substance solution: precisely weighing appropriate amount of hesperidin control, and quantitatively diluting with methanol to obtain solution containing 0.4mg per 1 ml.
(3) And (3) determination:
precisely measuring the sample solution and the reference solution by 10 μ l each, injecting into liquid chromatograph, measuring according to the above chromatographic conditions, and recording chromatogram. The content of hesperidin in the test solution is calculated by external standard method with reference solution.
According to the HPLC detection method for the dried orange peel component in the tortoise plastron and deer kidney tonifying capsule, the detector is an ultraviolet-visible light spectrum detector.
The HPLC detection method of the dried orange peel component in the tortoise plastron and deer kidney tonifying capsule detects the content of the dried orange peel component hesperidin in the tortoise plastron and deer kidney tonifying capsule by an HPLC method, separates the main quality control index hesperidin of dried orange peel from other components, and can independently determine the content of hesperidin.
The following is the methodology verification condition of the HPLC detection method of the dried orange peel component in the tortoise plastron and deer kidney tonifying capsule of the invention:
1. specificity test
Preparing a negative control solution: and preparing a negative control solution according to the prescription proportion and the preparation method of the test solution. The solvent, the negative control solution, the sample solution and the control solution were each precisely aspirated by 10. mu.l each, measured by the aforementioned detection method, and chromatograms were recorded (see FIG. 1, FIG. 2, FIG. 3 and FIG. 4). As can be seen from the figure, in the chromatogram of the test solution, both hesperidin and the adjacent peak can achieve baseline separation, and both the solvent and the negative control have no interference, which indicates that the method has good specificity.
2. Investigation of linear relationships
Accurately weighing 10.50mg of hesperidin reference substance, placing the hesperidin reference substance in a 10ml volumetric flask, adding methanol for dissolving, quantitatively diluting the hesperidin reference substance to a scale, shaking the volume evenly to obtain reference substance storage solution, accurately weighing a proper amount of the reference substance storage solution, quantitatively diluting the reference substance storage solution with methanol to obtain standard curve solutions with different concentrations of 1-7, taking the prepared standard curve solution, carrying out sample injection measurement, and carrying out linear regression on a peak area y by using a concentration x (mu g/ml) to obtain a result: the linear relation is good in the range of 51.37 mu g/ml to 1027.42 mu g/ml, the linear equation is that y is 18.513x-99.937, and the correlation coefficient r is 0.9994 (see figure 5).
3. Investigation of solution stability
A batch of samples (batch No. 190902) produced by Guangxi Huatianbao pharmaceutical industry Co., Ltd are taken to prepare a test sample solution, the test sample solution is respectively placed for 0 hour, 4 hours, 8 hours, 12 hours and 16 hours, then the sample injection is carried out for measurement, the result shows that the test sample solution is stable within 16 hours, the RSD of the hesperidin peak area is 0.45%, and the measurement result is shown in the following table 1.
TABLE 1 solution stability test
4. Sample introduction precision test
Taking a batch of samples (batch number 190902) produced by Guangxi Huatianbao pharmaceutical industry Co., Ltd to prepare 1 part of the test solution according to the preparation method of the test solution in the detection method, continuously feeding a sample of 6 needles, measuring the peak area of hesperidin, and inspecting the precision of the instrument, wherein the RSD of the peak area of hesperidin is 0.51 percent, which indicates that the precision of the instrument is good. The results of the measurements are shown in Table 2 below.
TABLE 2 sample introduction precision test
5. Repeatability test
A batch of samples (batch No. 190902) produced by Guangxi Huatianbao pharmaceutical industry Co., Ltd is taken, 6 parts of test solution is prepared according to the preparation method of the test solution in the detection method, the content of hesperidin and RSD are calculated by sample injection measurement, and the result is shown in the following table 3.
TABLE 3 repeatability tests
6. Sample application recovery test
Precisely weighing 22.36mg of hesperidin reference substance, placing the hesperidin reference substance in a 500ml volumetric flask, adding methanol for dissolving, quantitatively diluting to a scale, shaking uniformly to obtain a reference substance sample adding solution, precisely weighing 6 parts of a sample (a sample with the batch number of 190902, produced by Guangxi Huatianbao pharmaceutical industry Co., Ltd.) with the known content, placing 0.5g of each part in a conical flask with a plug, precisely weighing 50ml of the reference substance sample adding solution in the conical flask with the plug, preparing a solution with the recovery rate of 100% according to the preparation method of the sample solution in the detection method, and carrying out sample injection measurement, wherein the result is shown in the following table 4. According to the determination result, the HPLC detection method for the dried orange peel component in the tortoise plastron and deer kidney tonifying capsule is simple to operate and wide in applicability.
TABLE 4 sample recovery test
The invention has the advantages that
The HPLC detection method for the dried orange peel component in the tortoise and deer kidney tonifying capsule has the advantages of simple operation, high detection efficiency, small error, good reproducibility, strong specificity, stable detection method, fewer influenced factors and wide applicability; the specificity, stability, precision, accuracy and the like of the method all meet the requirement of methodology verification, and the method has strong practicability.
Drawings
FIG. 1 liquid chromatogram of solvent
FIG. 2 liquid chromatogram of negative control solution
FIG. 3 liquid chromatogram of test solution
FIG. 4 liquid chromatogram of control solution
FIG. 5 is a linear relationship graph of the hesperidin concentration x (μ g/ml) in the standard curve solution prepared by the present invention and the peak area y
Detailed Description
Example 1
An HPLC detection method for a dried orange peel component in a tortoise plastron and deer kidney tonifying capsule comprises the following steps:
(1) chromatographic conditions and system applicability test:
octadecylsilane chemically bonded silica is used as a filler, and a chromatographic column comprises the following components: waters
T3C 18 column, 4.6X 250mm, 5 μm or equivalent column, mobile phase: acetonitrile-water (22: 78). Flow rate: 1.0ml/min, detection wavelength: 283nm, column temperature: 35 ℃, sample introduction: 10 mul, the theoretical plate number is not lower than 5000 calculated according to the hesperidin peak.
(2) Solution preparation:
preparation of a test solution: taking the contents of the product with different filling amounts, mixing uniformly, grinding, taking about 1g, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of methanol, sealing the plug, weighing, heating and refluxing for 3 hours, cooling, weighing again, supplementing the lost weight with methanol, shaking up, filtering, precisely weighing 20ml of subsequent filtrate, recovering the solvent to dryness, adding a proper amount of methanol into the residue for dissolving, transferring to a 5ml measuring flask, adding methanol to scale, shaking up, filtering, taking the subsequent filtrate, centrifuging at 5000 r/min for 5min, and filtering with a 0.45 mu m filter membrane to obtain the sample solution.
Preparing a reference substance solution: precisely weighing appropriate amount of hesperidin control, and quantitatively diluting with methanol to obtain solution containing 0.4mg per 1 ml.
(3) And (3) determination:
precisely measuring the sample solution and the reference solution by 10 μ l each, injecting into liquid chromatograph, measuring according to the above chromatographic conditions, and recording chromatogram. The content of hesperidin in the test solution is calculated by external standard method with reference solution.
Therefore, the HPLC detection method for the dried orange peel component in the tortoise-deer kidney tonifying capsule has the advantages of simple operation, high detection efficiency, small error, good reproducibility and wide applicability; the method for detecting the dried orange peel component in the tortoise-deer kidney tonifying capsule can quickly and accurately detect the content of hesperidin, thereby being beneficial to judging the quality of the tortoise-deer kidney tonifying capsule.
Claims (3)
1. The HPLC detection method for the dried orange peel component in the tortoise-deer kidney tonifying capsule is characterized by comprising the following steps:
(1) chromatographic conditions and system applicability test:
octadecylsilane chemically bonded silica is used as a filler, and a chromatographic column comprises the following components: waters Atlantis
T3C 18 column, 4.6X 250mm, 5 μm or equivalent column, mobile phase: acetonitrile-water (22: 78). Flow rate: 1.0ml/min, detection wavelength: 283nm, column temperature: 35 ℃, sample introduction: 10 mul, the theoretical plate number is not lower than 5000 calculated according to the hesperidin peak.
(2) Solution preparation:
preparation of a test solution: taking the contents of the product with different filling amounts, mixing uniformly, grinding, taking about 1g, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of methanol, sealing the plug, weighing, heating and refluxing for 3 hours, cooling, weighing again, supplementing the lost weight with methanol, shaking up, filtering, precisely weighing 20ml of subsequent filtrate, recovering the solvent to dryness, adding a proper amount of methanol into the residue for dissolving, transferring to a 5ml measuring flask, adding methanol to scale, shaking up, filtering, taking the subsequent filtrate, centrifuging at 5000 r/min for 5min, and filtering with a 0.45 mu m filter membrane to obtain the sample solution.
Preparing a reference substance solution: precisely weighing appropriate amount of hesperidin control, and quantitatively diluting with methanol to obtain solution containing 0.4mg per 1 ml.
(3) And (3) determination:
precisely measuring the sample solution and the reference solution by 10 μ l each, injecting into liquid chromatograph, measuring according to the above chromatographic conditions, and recording chromatogram. The content of hesperidin in the test solution is calculated by external standard method with reference solution.
2. The HPLC detection method for the dried orange peel component in the tortoise-deer kidney tonifying capsule as claimed in claim 1, wherein the detector is an ultraviolet-visible light spectrum detector.
3. The HPLC detection method for the tangerine peel component in the tortoise-deer kidney tonifying capsule as claimed in claim 1, wherein the tangerine peel component hesperidin in the tortoise-deer kidney tonifying capsule is separated from other components by an HPLC method, and the content of hesperidin can be measured independently.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110445464.3A CN113916999A (en) | 2021-04-19 | 2021-04-19 | HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110445464.3A CN113916999A (en) | 2021-04-19 | 2021-04-19 | HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113916999A true CN113916999A (en) | 2022-01-11 |
Family
ID=79232734
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110445464.3A Withdrawn CN113916999A (en) | 2021-04-19 | 2021-04-19 | HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113916999A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1895617A (en) * | 2006-06-23 | 2007-01-17 | 贵阳德昌祥药业有限公司 | Kidney-warming and heart-nourishing Chinese-medicinal preparation, its making method and quality control |
| CN103285306A (en) * | 2013-06-09 | 2013-09-11 | 荣昌制药(淄博)有限公司 | Preparation method and detection method of traditional Chinese medicine composition for benefiting Qi and tonifying kidney |
| CN104062374A (en) * | 2014-06-27 | 2014-09-24 | 荣昌制药(淄博)有限公司 | Method for detecting traditional Chinese medicine composition for tonifying qi and tonifying kidney |
-
2021
- 2021-04-19 CN CN202110445464.3A patent/CN113916999A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1895617A (en) * | 2006-06-23 | 2007-01-17 | 贵阳德昌祥药业有限公司 | Kidney-warming and heart-nourishing Chinese-medicinal preparation, its making method and quality control |
| CN103285306A (en) * | 2013-06-09 | 2013-09-11 | 荣昌制药(淄博)有限公司 | Preparation method and detection method of traditional Chinese medicine composition for benefiting Qi and tonifying kidney |
| CN104062374A (en) * | 2014-06-27 | 2014-09-24 | 荣昌制药(淄博)有限公司 | Method for detecting traditional Chinese medicine composition for tonifying qi and tonifying kidney |
Non-Patent Citations (6)
| Title |
|---|
| 席艳;: "HPLC梯度洗脱联合波长切换法测定龟鹿补肾胶囊中7种成分的含量" * |
| 李文静;刘洁琼;李坤杰;秦峰;李发美;: "龟鹿补肾丸的质量控制" * |
| 李文静;洪博;: "RP-HPLC法同时测定龟鹿补肾丸中的4个主要活性成分" * |
| 李文静;洪博;: "龟鹿补肾丸HPLC指纹图谱研究" * |
| 王新建;覃建兵;刘会东;化宏伟;: "龟鹿补肾片质量标准研究" * |
| 钟涧: "高效液相色谱法测定龟鹿补肾胶囊中淫羊藿苷的含量" * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107796892B (en) | Fingerprint of Gentiana macrophylla capsule and its application in quality control and component analysis | |
| CN113125595A (en) | HPLC detection method of sophora flavescens component | |
| CN112098556A (en) | Detection method of angelica sinensis Liuhuang decoction | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN103197026A (en) | Quality control method of granules capable of strengthening and consolidating body resistance | |
| CN103822975A (en) | Test method for pulse-activating preparation adopting pilose asiabell root | |
| CN109900829B (en) | Quality control method of heat-clearing and cough-relieving granules | |
| CN110806457B (en) | Detection method of fingerprint of Sijun manna drink | |
| CN112730674A (en) | Quality detection method of momordica grosvenori tea | |
| CN110967422A (en) | Method for determining content of astragaloside in qi-tonifying and blood-nourishing tablet | |
| CN113916999A (en) | HPLC detection method for dried orange peel component in tortoise and deer kidney tonifying capsule | |
| CN106442748B (en) | Content determination method of liver-soothing pills | |
| CN111351883B (en) | Method for measuring rutin content in Sophora japonica and radix scutellariae ointment | |
| CN111929378B (en) | Method for measuring content of 6 index components of gastrodia elata in Qingda granules | |
| CN114563496A (en) | Quantitative fingerprint analysis method for components in ginger, ginger and pinellia tuber percolate | |
| CN1947740B (en) | Lamiophlomis rotata medicine material, intermediate and its injection liquid finger-print atlas quality testing method | |
| CN103575823A (en) | Detection method of 8 chemical components in Tangminling preparation | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN101669977A (en) | Quality detection method of dogbane leaf capsule | |
| CN113960234B (en) | Quality control method for cynanchum glaucescens and cynanchum glaucescens formula granules | |
| CN108627599A (en) | A kind of Cortex Eucommiae is the content assaying method that raw material is prepared into drug containing Cortex Eucommiae | |
| CN114894922B (en) | Detection and quality control method of gynostemma pentaphylla water extract | |
| CN102539562A (en) | Detection method for kidney tonifying and vigour nourishing mixture | |
| CN114791468B (en) | Detection method of mountain branch kernel medicinal material, decoction piece, extract and preparation | |
| CN115372516B (en) | Method for measuring content of nucleoside components in houttuynia cordata, radix scutellariae and blue mixture intermediate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220111 |
|
| WW01 | Invention patent application withdrawn after publication |