CN114487241B - Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof - Google Patents
Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof Download PDFInfo
- Publication number
- CN114487241B CN114487241B CN202111619451.XA CN202111619451A CN114487241B CN 114487241 B CN114487241 B CN 114487241B CN 202111619451 A CN202111619451 A CN 202111619451A CN 114487241 B CN114487241 B CN 114487241B
- Authority
- CN
- China
- Prior art keywords
- detection
- tortoise
- performance liquid
- high performance
- liquid chromatography
- 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.)
- Active
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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
-
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for detecting tortoise shell, vinegar tortoise shell and their formula granules and quality control. The detection method provided by the invention comprises the following steps: obtaining a test solution of an object to be tested, wherein the preparation method of the test solution comprises the following steps: obtaining a substance to be measured, precisely weighing, adding a solvent, sealing, weighing, ultrasonically treating, cooling, weighing again, complementing the lost weight with the corresponding solvent, shaking up, filtering, and taking a subsequent filtrate; directly carrying out high performance liquid chromatography detection on the test solution to obtain a characteristic spectrum; methanol is used as a mobile phase A and 0.08-0.12% phosphoric acid aqueous solution is used as a mobile phase B in the high performance liquid chromatography detection. The invention solves the problems of complex preparation, high cost and environmental pollution of the test solution in the existing high performance liquid chromatography for detecting tortoise shells, vinegar tortoise shells and formula granules thereof; the quick and simple detection of the tortoise shell, the vinegar tortoise shell and the formula granules thereof can be effectively realized under the condition of not needing acidolysis and derivative reagents.
Description
Technical Field
The invention relates to the field of traditional Chinese medicine detection, in particular to a method for detecting tortoise shells, vinegar tortoise shells and formula granules thereof and controlling the quality of the tortoise shells and the vinegar tortoise shells.
Background
The carapax et Plastrum Testudinis is the dorsal and ventral concha of Chinemys reevesii (Gray) of Testudinidae. It has effects in nourishing yin, suppressing yang hyperactivity, invigorating kidney, strengthening bone, nourishing blood, tonifying heart, consolidating channels, and relieving metrorrhagia, and can be used for treating hectic fever due to yin deficiency, night sweat due to bone steaming, dizziness, blurred vision, internal stirring due to deficient wind, tendons and bones flaccidity, heart deficiency, amnesia, and metrorrhagia. According to the examination, tortoise shells are listed as the superior product from Shen nong Ben Cao Jing, which is a common traditional Chinese medicine in clinical practice of traditional Chinese medicine. The processed tortoise shell with vinegar has the same functions and indications, but has strengthened Yin nourishing effect and can correct odor and taste.
At present, the high performance liquid chromatography-mass spectrometry combined technology is adopted for the ingredient research of tortoise shells, or the high performance liquid chromatography is directly adopted. Wherein, when the high performance liquid chromatography-mass spectrometry combined technology is adopted, the test sample is generally detected after enzymolysis; in each local formula particle quality standard, for example: BJ-PFKL-2021046 discloses that a test sample is treated by ammonium bicarbonate solution and then is subjected to enzymolysis by trypsin solution to prepare a test sample solution meeting the detection requirement of a high performance liquid chromatography-mass spectrometry combined technology.
When the high performance liquid chromatography is directly adopted, the test sample needs to be firstly subjected to acidolysis by adopting an acid solution and then subjected to derivatization by adopting a derivatization reagent; for example: in the existing documents, "HPLC fingerprint spectrum research of tortoise shell as a traditional Chinese medicine, lv Xiaona and the like", "determination of content of 6 amino acids in tortoise shell extract by using one-test-multiple evaluation method, zhou Wei and the like", acidic reagents such as acetic acid, hydrochloric acid and the like are firstly adopted to carry out acidolysis on a sample, and then derivatization treatment is carried out by adopting derivatization reagents such as Phenyl Isothiocyanate (PITC), phthalaldehyde (OPA), 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate (AQC) and the like. In the preparation process of the test solution, a derivatization reagent is a necessary condition, and the preparation method is relatively complicated, time-consuming, high in cost, not environment-friendly and not beneficial to wide popularization and application.
In addition, both the high performance liquid chromatography-mass spectrometry technology and the high performance liquid chromatography are characterized by amino acid components, amino acid widely exists in various substances in the nature, and the specificity of the characterization on the specific substances of the tortoise shells is not strong.
Disclosure of Invention
Therefore, the invention solves the problems that the preparation of a test solution is complex, the cost is high and the environment is not protected in the conventional high performance liquid chromatography for detecting the tortoise shells, the vinegar tortoise shells and the formula particles thereof, and provides the detection method which can effectively realize the detection of the tortoise shells, the vinegar tortoise shells and the formula particles thereof without acidolysis and derivative reagents.
A method for detecting tortoise shells, vinegar tortoise shells and formula granules thereof comprises the following steps:
obtaining a test solution of an object to be tested, wherein the preparation method of the test solution comprises the following steps: obtaining a substance to be measured, precisely weighing, adding water, sealing, weighing, ultrasonically treating, cooling, weighing again, supplementing the lost weight with water, shaking up, filtering, and taking a subsequent filtrate;
directly carrying out high performance liquid chromatography detection on the test solution to obtain a characteristic spectrum; methanol is used as a mobile phase A, and 0.08-0.12% phosphoric acid aqueous solution is used as a mobile phase B in the high performance liquid chromatography detection.
The chromatographic conditions of the high performance liquid chromatography detection are as follows: chromatographic column with octadecyl bonded silica gel as filler; detection wavelength: 210-230nm; elution was performed according to the following gradient elution procedure:
the column temperature of the chromatographic column in the high performance liquid chromatography is 29-31 ℃, the flow rate is 0.75-0.85ml/min, and the number of theoretical plates is not less than 5000 according to the tyrosine peak.
The chromatographic column adopts an Xselect HSS T3 chromatographic column.
The gradient elution procedure was:
the to-be-detected substances are tortoise shell, vinegar tortoise shell, tortoise shell aqueous extract, vinegar tortoise shell aqueous extract, tortoise shell formula particles and vinegar tortoise shell formula particles.
The power of the ultrasonic wave is 250W, the frequency is 40kHz, and the time is 30min.
A quality control method for tortoise shell, vinegar tortoise shell and their formula granule is characterized by that it adopts the above-mentioned detection method to detect and obtain characteristic spectrum, in the described characteristic spectrum at least includes 6 characteristic peaks, the peak in which tyrosine is positioned is S peak, the relative retention time of other characteristic peaks is in the range of plus or minus 10% of defined value, and the defined value of the relative retention time of every characteristic peak is: 0.20, 0.50, 0.57, 0.72, 0.76, 1.00.
The technical scheme of the invention has the following advantages:
1. the invention provides a detection method for tortoise shells, vinegar tortoise shells and formula particles thereof, wherein the chromatographic conditions are optimized, specifically, methanol is used as a mobile phase A,0.08-0.12% phosphoric acid aqueous solution is used as a mobile phase B, at the moment, a to-be-detected object can be prepared into a test solution only by adopting water for ultrasonic treatment, the test solution is obtained without acid hydrolysis of an acidic solution or derivatization of a derivatization reagent, the treatment conditions are simple, the treatment cost is low, and the method is environment-friendly, pollution-free and very convenient to popularize and apply.
2. The high performance liquid chromatography provided by the invention can be used for simultaneously detecting the tortoise shell and vinegar tortoise shell formula particles, is stable, high in precision and better in reproducibility, and provides a rapid and comprehensive detection means for the quality of the tortoise shell and vinegar tortoise shell formula particles.
3. The quality control method of the tortoise shell and vinegar tortoise shell and the formula particles thereof provided by the invention has the characteristic peaks that not only contain amino acid components, but also contain exclusive components of the tortoise shell and vinegar tortoise shell which are not amino acid, and can comprehensively reflect the characteristic peak information of a sample; therefore, the quality control method of the invention can strengthen specificity identification and integral quality control, and can realize authenticity or quality identification of tortoise shells and vinegar tortoise shells.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
Fig. 1 is a characteristic map of three batches of tortoise shell formula granules in example 1 of the present invention.
Fig. 2 is a comparison feature map obtained by fitting chromatograms of three batches of tortoise shell formula particles in example 1 of the present invention.
Fig. 3 is a comparative profile of tortoise shell formulation granule and reference solution in example 1 of the present invention.
Fig. 4 is a comparative characteristic spectrum of the tortoise-shell formulation granule, the vinegar tortoise-shell formulation granule and the control solution in example 1 of the present invention.
FIG. 5 is a comparative profile for different columns in example 2 of the invention.
FIG. 6 is a characteristic diagram of the wavelength of 210nm in example 2 of the present invention.
FIG. 7 is a characteristic diagram of the wavelength of 220nm in example 2 of the present invention.
FIG. 8 is a characteristic diagram of test solutions prepared by different solvents in example 2 of the present invention.
Detailed Description
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
The instrument comprises the following steps:
shimadzu 20AT high performance liquid chromatograph: PDA detector, labsolutions chromatography workstation; waters 2695 high performance liquid chromatograph: UV detector, empower 3 chromatography workstation; XP26 (mettler-toledo), BSA124S electronic balance (sydows scientific instrument (beijing) ltd), BT25S electronic balance (sydows scientific instrument (beijing) ltd), KQ-500DB ultrasonic cleaner (kunshan ultrasonic instrument ltd).
Reagent testing:
tortoise shell formula particle (batch number: 19011411, 19011372, 20021681)
Vinegar tortoise shell formula particle (batch number: 19036391)
Tortoise shell reference medicine (batch number: 121494-201604, china institute for testing food and drug)
Tyrosine (batch number: 140609-201914, purity 99.9%, china institute for testing food and drug)
Reagent: methanol (Merck Co., ltd., JA 078530), phosphoric acid (Fisher Scientific, 171289) as chromatographically pure; the water is distilled water (drochen); other reagents were analytically pure.
Example 1
A method for detecting tortoise shells, vinegar tortoise shells and formula granules thereof comprises the following steps:
obtaining an object to be tested and preparing a test solution, wherein the object to be tested comprises three batches of tortoise shell formula particles and one batch of vinegar tortoise shell formula particles; the preparation process of the test solution comprises the following steps: taking about 0.5g of formula particles, accurately weighing, placing in a conical flask with a plug, accurately adding 25ml of water, sealing the plug, weighing, carrying out ultrasonic treatment (with the power of 250W and the frequency of 40 kHz) for 30 minutes, cooling, weighing again, supplementing the lost weight with water, shaking up, filtering, and taking the subsequent filtrate to obtain the test solution of the formula particles of tortoise shells and vinegar tortoise shells.
Preparing reference solution, wherein the reference solution comprises reference medicinal material solution and reference substance solution. Wherein, the preparation process of the reference medicinal material solution comprises the following steps: collecting carapax et Plastrum Testudinis reference medicinal material about 1.0g, adding water 25ml, decocting for 40 min, filtering, and collecting filtrate; the preparation process of the reference solution comprises the following steps: taking appropriate amount of tyrosine reference substance, precisely weighing, and adding 30% methanol to obtain solution containing 0.01mg per 1 ml.
And (3) obtaining the characteristic maps of the test solution and the reference solution by adopting a high performance liquid chromatography.
Specifically, the chromatographic conditions of the high performance liquid chromatography in this example are: octadecylsilane chemically bonded silica was used as a filler (column length 25cm, inner diameter 4.6mm, particle diameter 5 μm); gradient elution was performed with methanol as mobile phase a and 0.1% phosphoric acid aqueous solution as mobile phase B as specified in table 1 below; the flow rate was 0.8ml per minute; the detection wavelength is 230nm; the column temperature is 30 ℃, and the number of theoretical plates is not less than 5000 according to the tyrosine peak.
TABLE 1
The determination method comprises the following steps: precisely absorbing 5 μ l of reference solution and 10 μ l of test solution, respectively, injecting into high performance liquid chromatograph, measuring, introducing the chromatogram into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system for data matching, and establishing reference characteristic chromatogram of carapax et Plastrum Testudinis formula granule, with the results shown in figures 1-4. Wherein, fig. 1 is a characteristic map of three batches of tortoise shell formula particles, fig. 2 is a comparison characteristic map obtained after fitting chromatograms of three batches of tortoise shell formula particles, fig. 3 is a comparison characteristic map of one batch of tortoise shell formula particles and a reference substance solution, and fig. 4 is a comparison characteristic map of tortoise shell formula particles, vinegar tortoise shell formula particles and a reference substance solution.
Example 2
The difference between the detection method for tortoise shells, vinegar tortoise shells and formula granules thereof and the detection method for the tortoise shells and the vinegar tortoise shells in the embodiment 1 is that the chromatographic conditions are different, and the specific settings are as follows:
1. detection of different column temperatures
In this example, characteristic spectrum detection was performed on the test solution of the tortoise shell formula particle according to the method of example 1 at 29 ℃, 30 ℃ and 31 ℃, and the detection results are shown in table 2 below.
TABLE 2 relative retention time tables at different column temperatures
And (3) knotting: from the above data, it can be seen that the relative retention times of the 6 peaks of the signatures obtained at the other two temperatures are within + -10% of the relative retention times of the signatures obtained at 30 ℃, indicating that the method is well-tolerated at column temperatures between 29 ℃ and 31 ℃.
2. Detection of different acid concentrations
In this example, the concentration of the phosphoric acid solution is 0.08%, 0.10%, 0.12%, and the characteristic spectrum of the test solution of the tortoise shell formula granule is determined according to the method of example 1, and the results are shown in table 3 below.
TABLE 3 relative Retention time Table for different acid concentrations
And (3) knotting: from the above data, it can be seen that the relative retention time of the 6 peaks is within ± 10% of the specified value, and the chromatographic peak separation is better at different acid concentrations, indicating that a concentration of the phosphoric acid solution in the range of 0.08-0.12% has less influence on the process.
3. Differential flow rate detection
In this example, the characteristic spectrum detection of the test solution of the tortoise shell formula particle is performed according to the method of example 1 at the flow rates of 0.75ml/min, 0.80ml/min and 0.85ml/min, respectively, and the detection results are shown in table 4 below.
TABLE 4 relative Retention time tables at different flow rates
And (3) knotting: from the above data, it can be seen that the relative retention time of the 6 peaks is within + -10% of the specified value, and the chromatographic peak separation is better at different flow rates, indicating that flow rates in the range of 0.75-0.85ml/min have less effect on the process.
4. Detection of different chromatographic columns
In this example, xbridge Shield RP18 and WondaSil C18 chromatography columns were used for detection, and other characteristic maps of the test solution of the tortoise shell formulation granule were performed according to the method of example 1, and the detection results are shown in fig. 5 below. As can be seen from fig. 5, the peak shape and the separation of the columns of different brands are greatly different, and the Xselect HSS T3 column is preferred in this embodiment.
5. Detection of different wavelengths
In this example, the wavelength 210nm and the wavelength 220nm are respectively detected, and the characteristic spectrum detection is performed on the test solution of the tortoise shell formula particle according to the method in example 1, and the detection results are shown in table 5 below.
TABLE 5
As can be seen from Table 5 above and FIGS. 6 and 7, the spectral peaks at different wavelengths are well separated, indicating that wavelengths in the range of 210-230nm have less effect on the process.
6. Detection of different solvents
The preparation method comprises the steps of taking tortoise shell particles, grinding, taking 6 parts of the tortoise shell particles, precisely weighing, placing the tortoise shell particles in conical bottles with plugs, precisely adding 10% methanol aqueous solution, 30% methanol aqueous solution, 50% methanol aqueous solution, 70% methanol aqueous solution, methanol and water with volume concentration of 25ml, sealing, weighing, carrying out ultrasonic treatment (power of 250W and frequency of 40 kHz) for 30 minutes, taking out, cooling, weighing again, supplementing the reduced weight with corresponding solvents respectively, shaking up, filtering, precisely sucking 10 mu l of subsequent filtrate, injecting into a high performance liquid chromatograph, measuring according to determined chromatographic conditions, and obtaining a result shown in a figure 8.
And (3) knotting: from the above results, it can be seen that the peak shape and the resolution of the obtained pattern are good when an alcohol solution having a volume concentration of 50% or less and water are used as the extraction solvent, and that water is preferred as the extraction solvent because the peak shape and the resolution of the pattern when water is used as the extraction solvent are superior to those of the alcohol solution.
Example 3
This example is used to verify the precision and stability of the detection method for tortoise shells, vinegar tortoise shells and their formula granules described in example 1, and specifically includes the following steps:
1. precision degree
1.1, reproducibility
6 parts of tortoise shell formula particles (batch No. 19011411) are taken, the characteristic spectrum is obtained by measuring according to the method described in example 1, and the relative peak area and the relative retention time are calculated by taking the No. 6 peak as a reference peak. And calculates the RSD. The results are shown in tables 6 and 7.
TABLE 6 retention time and relative retention time table for repeated survey of feature profiles
TABLE 7 characteristic chromatogram repeatability survey peak area and relative peak area
And (4) summarizing: according to the repeatability inspection result, the relative retention time RSD of each characteristic peak is 0% -0.3%, and the RSD of the relative peak area is in the range of 0% -2.3%, which shows that the repeatability of the characteristic spectrum is good.
1.2, intermediate precision
6 parts of tortoise shell formula particles (batch number: 19011411) are taken by a Waters 2695 UV detector, a characteristic spectrum is obtained by measuring according to the method of the technical scheme, and the relative peak area and the relative retention time are calculated by taking the No. 6 peak as a reference peak. And calculates the RSD. The results are shown in tables 8 and 9.
TABLE 8 intermediate precision survey of retention times and relative retention time tables for feature maps
TABLE 9 middle precision survey peak area and relative peak area table of characteristic spectrum
And (4) summarizing: in the characteristic spectrum obtained by using Waters 2695, the relative retention time RSD of each characteristic peak is in the range of 0-0.6%, and the RSD of the relative peak area is in the range of 0-2.3%. The relative retention time RSD range of different instruments is 0-9.4%, and the relative peak area RSD% range is 0-81.0%, which shows that the analysis of different brands of instruments has certain influence on the characteristic spectrum.
2. Stability of solution
Taking the test solution of the tortoise shell formula particle, respectively measuring in 0,2,4,6,8,10, 12 and 24h, analyzing characteristic peaks in the test solution, wherein the relative retention time of corresponding chromatographic peaks is less than 1.0%, and the RSD of the relative peak area of the characteristic peaks is less than 3.0%, and the result shows that the test solution is stable in 24 h. See tables 10 and 11.
TABLE 10 stability relative Retention time Table
TABLE 11 table of stability versus peak area
And (3) knotting: the stability experiment result shows that the chemical components in the test solution are stable in 24 hours, the RSD of the relative retention time of each characteristic peak is within 0-0.5%, and the RSD of the relative peak area is within 0-2.9%, so that the analysis method is stable and reliable.
Example 4
A quality control method for carapax et Plastrum Testudinis, carapax et Plastrum Testudinis and vinegar carapax et Plastrum Testudinis and its formula granule comprises displaying 6 characteristic peaks in characteristic chromatogram of carapax et Plastrum Testudinis formula granule, corresponding to 6 characteristic peaks retention time in reference chromatogram peak of carapax et Plastrum Testudinis reference material, wherein peak 6 corresponds to tyrosine; calculating the relative retention time of each characteristic peak and the S peak by taking the peak corresponding to the tyrosine reference substance peak as the S peak, wherein the relative retention time is within +/-10% of a specified value: 0.20 (peak 1), 0.50 (peak 2), 0.57 (peak 3), 0.72 (peak 4), 0.76 (peak 5), 1.00 (peak 6).
In this example, the characteristic spectrum of 3 batches of tortoise shell formula granules was obtained by the detection method described in example 1, and the detection results of the 3 batches of tortoise shell formula granules are shown in table 12 below.
TABLE 12 batch Tortoise Shell granule relative Retention time Table
As can be seen from the characteristic maps of the 3 batches of tortoise shell particles, the relative retention time difference of each characteristic peak is small and is within the range of +/-10%, thus meeting the quality control requirements.
And as can be seen from the detection result of fig. 4 in example 1, the characteristic peaks of the particle spectra of the tortoise shell and the vinegar tortoise shell correspond well, so that the method can quickly and comprehensively detect the formula particles of the tortoise shell and the vinegar tortoise shell, and provides a more scientific basis for the quality control of the formula particles of the tortoise shell and the vinegar tortoise shell.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (5)
1. A detection method for water extracts and formula granules of tortoise shells and vinegar tortoise shells is characterized by comprising the following steps:
obtaining a test solution of an object to be tested, wherein the preparation method of the test solution comprises the following steps: obtaining a substance to be measured, precisely weighing, adding a solvent, sealing, weighing, ultrasonically treating, cooling, weighing again, complementing the lost weight with the corresponding solvent, shaking up, filtering, and taking a subsequent filtrate;
directly carrying out high performance liquid chromatography detection on the test solution to obtain a characteristic spectrum; in the high performance liquid chromatography detection, methanol is used as a mobile phase A, and 0.08-0.12% phosphoric acid aqueous solution is used as a mobile phase B; the solvent is an alcohol solution or water with the volume concentration of below 50%; the chromatographic conditions of the high performance liquid chromatography detection are as follows: an Xselect HSS T3 chromatographic column is adopted; detection wavelength: 210-230nm; elution was performed according to the following gradient elution procedure:
。
2. The detection method according to claim 1, wherein the column temperature of the chromatographic column in the high performance liquid chromatography is 29-31 ℃, the flow rate is 0.75-0.85ml/min, and the number of theoretical plates is not less than 5000 calculated according to tyrosine peak.
3. The detection method according to claim 1 or 2, wherein the gradient elution procedure is replaceable by:
。
4. The detection method according to claim 1, wherein the power of the ultrasound is 250W, the frequency is 40kHz, and the time is 30min.
5. A quality control method for water extracts and formula granules of tortoise shell and vinegar tortoise shell and both of them is characterized in that a characteristic spectrum is obtained by the detection method of any one of claims 1 to 4, the characteristic spectrum at least comprises 6 characteristic peaks, wherein the peak of tyrosine is the S peak, the relative retention time of the rest characteristic peaks is within +/-10% of a specified value, and the specified value of the relative retention time of each characteristic peak is as follows: 0.20, 0.50, 0.57, 0.72, 0.76, 1.00.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111619451.XA CN114487241B (en) | 2021-12-27 | 2021-12-27 | Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111619451.XA CN114487241B (en) | 2021-12-27 | 2021-12-27 | Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114487241A CN114487241A (en) | 2022-05-13 |
| CN114487241B true CN114487241B (en) | 2023-03-28 |
Family
ID=81495361
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111619451.XA Active CN114487241B (en) | 2021-12-27 | 2021-12-27 | Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114487241B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527362A (en) * | 2015-12-17 | 2016-04-27 | 天津市天津医院 | Analysis method of fingerprint spectrum of amino acids in granulation-promoting ointment |
| CN113655151A (en) * | 2021-09-07 | 2021-11-16 | 广东一方制药有限公司 | Animal traditional Chinese medicine standard decoction amino acid characteristic map construction and traditional Chinese medicine standard decoction and traditional Chinese medicine formula granule amino acid content detection |
-
2021
- 2021-12-27 CN CN202111619451.XA patent/CN114487241B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527362A (en) * | 2015-12-17 | 2016-04-27 | 天津市天津医院 | Analysis method of fingerprint spectrum of amino acids in granulation-promoting ointment |
| CN113655151A (en) * | 2021-09-07 | 2021-11-16 | 广东一方制药有限公司 | Animal traditional Chinese medicine standard decoction amino acid characteristic map construction and traditional Chinese medicine standard decoction and traditional Chinese medicine formula granule amino acid content detection |
Non-Patent Citations (4)
| Title |
|---|
| 于海英 等.阿胶、龟甲胶中脂溶性成分的高效液相色谱指纹图谱.色谱.2009,第27卷(第4期),447-452. * |
| 吕晓娜 等.中药龟甲HPLC指纹图谱研究.世界中医药.2019,第14卷(第2期),311-314. * |
| 周维 等.一测多评法测定龟甲提取物中6种氨基酸的含量.现代中药研究与实践.2016,第30卷(第6期),57-61. * |
| 郭尚伟 等.HPLC-MS/MS法测定阿胶、龟甲胶、鹿角胶中17种氨基酸含量.明胶科学与技术.2016,第36卷(第2期),86-91. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114487241A (en) | 2022-05-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110286186B (en) | Method for detecting fingerprint of Xinhuang tablets, Xinhuang tablet standard fingerprint and application | |
| CN106018602B (en) | Combine detection reagent and its detection method for detecting sulfonamides compound | |
| CN114778731B (en) | Construction method and application of UPLC characteristic spectrum of endothelium corneum Gigeriae Galli, parched endothelium corneum Gigeriae Galli, vinegar endothelium corneum Gigeriae Galli decoction pieces, and standard soup and granule thereof | |
| CN114487241B (en) | Detection and quality control method for tortoise shells, vinegar tortoise shells and formula granules thereof | |
| CN113759037B (en) | Characteristic spectrum of formula granules of semen lepidii and/or semen lepidii as well as construction method and identification method thereof | |
| CN111855867B (en) | Method for establishing characteristic spectrum of traditional Chinese medicine or traditional Chinese medicine composition preparation and application | |
| CN108445115B (en) | Method for simultaneously detecting Niaoling, Songguoling and fuziline by high performance liquid chromatography | |
| CN114942291B (en) | Quality detection method of yang-suppressing yin-nourishing particles | |
| CN114216980B (en) | Method for establishing HPLC-ELSD fingerprint of starwort root | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN110441443B (en) | UPLC characteristic spectrum construction method and identification method of pyrrosia peduncularis, pyrrosia lingua, pyrrosia cottonii and pyrrosia huabeiensis | |
| CN110487938B (en) | Quality control method of podocarpus macrophyllus kurz medicinal material | |
| CN110361459B (en) | Method for establishing loquat leaf standard decoction fingerprint | |
| CN108982731B (en) | Method for constructing HPLC characteristic spectrum of sarcandra glabra buccal tablet and HPLC characteristic spectrum thereof | |
| CN107807182B (en) | Method for measuring content of ganoderic acid A in ganoderma lucidum syrup | |
| CN112858531A (en) | Method for establishing HPLC (high performance liquid chromatography) characteristic spectrum of eucommia leaves and extract thereof | |
| CN115372510B (en) | Hypocrellin fingerprint and establishment method and application thereof | |
| CN111487351A (en) | Method for detecting fingerprint of blood-activating pain-relieving capsule | |
| CN111505156A (en) | Fingerprint spectrogram quality determination method for herba Cirsii formulation granules | |
| CN114216978B (en) | Construction method of fingerprint of lindley eupatorium herb formula particle, standard fingerprint and application thereof | |
| CN112326806B (en) | Albizzia julibrissin formula particle HPLC characteristic spectrum and construction method and application thereof | |
| CN111766309B (en) | Detection method of Anshenbao particle characteristic spectrum | |
| CN113960234B (en) | Quality control method for cynanchum glaucescens and cynanchum glaucescens formula granules | |
| CN109374771A (en) | The fingerprint atlas detection method of snow pear paste | |
| CN112881541B (en) | Detection method of north bupleurum and south bupleurum prescription granule |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |