CN114487239A - Method for detecting fingerprint spectrum of instant momordica grosvenori solid beverage - Google Patents
Method for detecting fingerprint spectrum of instant momordica grosvenori solid beverage Download PDFInfo
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- CN114487239A CN114487239A CN202110877009.0A CN202110877009A CN114487239A CN 114487239 A CN114487239 A CN 114487239A CN 202110877009 A CN202110877009 A CN 202110877009A CN 114487239 A CN114487239 A CN 114487239A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
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- G—PHYSICS
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- 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
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- G—PHYSICS
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Abstract
The invention discloses a method for detecting a fingerprint of instant momordica grosvenori solid beverage, which comprises the following steps of 1, preparing a test solution; step 2, preparation of a mixed reference solution: step 3, respectively and precisely sucking the mixed reference solution and the test solution to be injected into a liquid chromatograph, and recording a chromatogram; step 4, leading out the fingerprint instrument of the instant momordica grosvenori solid beverage, leading the fingerprint instrument into a traditional Chinese medicine chromatography fingerprint similarity evaluation system, and selecting chromatographic peaks existing in the chromatograms of different batches of instant momordica grosvenori solid beverages as common peaks; generating a comparison fingerprint of the instant momordica grosvenori solid beverage by using an average value calculation method; calculating the relative retention time and the relative peak area of each common peak; and comparing the mixed standard substance spectra to identify the main component peak. The fingerprint spectrum established by the invention can comprehensively and objectively characterize the quality of the instant momordica grosvenori solid beverage. And the detection method has the advantages of simple and stable method, high precision, good reproducibility and the like.
Description
Technical Field
The invention relates to a detection method of a traditional Chinese medicine instant solid beverage, in particular to a detection method of a fingerprint spectrum of an instant momordica grosvenori solid beverage.
Background
With the development of industrialization, air pollution is serious, and the lung of a human body is threatened greatly. In addition, respiratory diseases also exacerbate lung injury. Therefore, the key of preventing virus infection and clearing lung pollution is to adjust the functions of respiratory tract and lung, and the invention starts with clearing lung, not only clears lung-heat and phlegm-heat stagnates fire, but also has the effects of nourishing yin and restoring yang, simultaneously enhances the immune function of the organism and resists virus invasion.
The momordica grosvenori is sweet, cool, clear and moist, and is a good product for clearing lung heat, relieving cough and moistening dryness as a main medicinal material; in the formula, the chrysanthemum is sweet and bitter, has cool property and can clear lung heat and be used for treating cough caused by lung heat; platycodon grandiflorum, being bitter in taste and neutral in nature, emphasizes on clearing lung and eliminating phlegm; the mulberry leaf, being bitter in taste and cold in nature, can dispel wind and heat, clear lung heat and moisten dryness, and has the effects of clearing away pathogenic heat in lung channels, ventilating lung and depressing qi, and reducing phlegm and relieving cough by combining the mulberry leaf, the mulberry leaf and the lung heat; the coix seed, the orange peel and the liquorice are all products for replenishing qi, eliminating dampness and strengthening spleen, and the spleen and stomach are healthy, so that the source of phlegm generation can be avoided. The gardenia and the dandelion can remove toxins generated by the lung, and discharge harmful substances through the functions of clearing heat and promoting the production of body fluid, relieving restlessness, quenching thirst and promoting urination of the reed rhizome and the lophatherum gracile; the dried ginger warms the middle-jiao to dispel cold, returns yang to dredge collaterals, and the ginseng warms the middle-jiao to tonify qi, strengthens the immune function of cells and body fluid and strengthens the virus resistance of the lung.
The components in the formula play their own roles, so that the functions of clearing heat, moistening lung, relieving sore throat, relieving cough, strengthening spleen and reducing phlegm are achieved, and the resistance of a human body is enhanced, so that the health-care food can be used for auxiliary food therapy of cough and profuse sputum due to phlegm-heat lung, and simultaneously, the health-care food can clear away pollutants in the lung and respiratory tract of the human body, protect the respiratory system, and have a good improvement effect on lung diseases caused by smoke, air pollution and the like; the deposited tar, nicotine and air pollutants are decomposed, and the decomposed harmful substances are discharged out of the body along with phlegm and urine.
At present, the quality detection methods of the instant momordica grosvenori solid beverage are few. The invention adopts the high performance liquid chromatography to establish the fingerprint detection method of the instant momordica grosvenori solid beverage, and has important significance for ingredient identification, quality evaluation and quality standard formulation of the instant momordica grosvenori solid beverage.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to solve the defects of the prior art and provides the fingerprint detection method of the instant momordica grosvenori solid beverage, and the detection method can objectively, comprehensively and accurately evaluate the quality of the instant momordica grosvenori solid beverage and has important significance in controlling the quality of the instant momordica grosvenori solid beverage and ensuring the clinical curative effect.
The technical scheme is as follows: in order to achieve the purpose, the invention adopts the technical scheme that:
a method for detecting the fingerprint spectrum of instant momordica grosvenori solid beverage is characterized by comprising the following steps:
accurately weighing different batches of instant fructus Siraitiae Grosvenorii solid beverage fluid extract samples, placing in round-bottomed flask, ultrasonic extracting, centrifuging extractive solution, and filtering supernatant with 0.45 μm microporous membrane to obtain sample solution;
precisely weighing chlorogenic acid and hesperidin as reference substances, placing in a volumetric flask, adding methanol to desired volume to scale, shaking to obtain mixed reference substance solution;
step 4, exporting the fingerprint of the instant momordica grosvenori solid beverage test solution obtained in the step 3, and importing the fingerprint into a traditional Chinese medicine chromatography fingerprint similarity evaluation system 2012A; selecting chromatographic peaks existing in chromatograms of different batches of instant siraitia grosvenorii solid beverages as common peaks; generating a comparison fingerprint of the instant momordica grosvenori solid beverage by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; marking chemical components of peaks in the comparison fingerprint spectrum according to the retention time of the mixed reference solution chromatogram;
and 5, comparing the fingerprint spectrum of the instant momordica grosvenori solid beverage obtained in the step 3 with the spectrum of the mixed standard substance, and identifying that the peak 6 in the instant momordica grosvenori solid beverage is chlorogenic acid and the peak 10 in the instant momordica grosvenori solid beverage is hesperidin.
As a preferred scheme, the method for detecting the fingerprint spectrum of the instant momordica grosvenori solid beverage comprises the following steps of 1: taking 5g of 8 batches of instant fructus momordicae solid beverage clear paste samples, placing the samples in a round bottom flask, adding 50ml of methanol, carrying out ultrasonic extraction for 30min, carrying out 12000-rotation centrifugation for 5min, and passing supernate through a 0.45 mu m microporous filter membrane to obtain a test solution.
Preferably, the method for detecting the fingerprint of the instant momordica grosvenori solid beverage comprises the following steps of 2, preparing a mixed reference solution: precisely weighing chlorogenic acid and hesperidin, placing in a volumetric flask, adding methanol to desired volume to scale, shaking, and making into mixed reference solution containing 0.1019mg/mL chlorogenic acid and 0.1101mg/mL hesperidin.
Preferably, in the method for detecting the fingerprint of the instant momordica grosvenori solid beverage, in step 3, the liquid chromatography conditions are as follows: a chromatographic column: YMC-Pack ODS-A, mobile phase: acetonitrile and 0.2% phosphoric acid water, gradient elution, diode array detector, detection wavelength: 210nm, column temperature 30 ℃, flow rate 0.6mL/min, sample injection volume: 10 μ L, gradient elution procedure as follows:
| Procedure | time (min) | Acetonitrile volume (%) |
| 1 | 0 | 5 |
| 2 | 15 | 5 |
| 3 | 65 | 21 |
| 4 | 85 | 21 |
| 5 | 95 | 22 |
| 6 | 110 | 40 |
As a preferable scheme, the fingerprint spectrum of the instant momordica grosvenori solid beverage detection method has 14 peaks in total in the fingerprint spectrum.
The instant momordica grosvenori solid beverage comprises the following medicinal components in parts by weight: 15-30 parts of momordica grosvenori, 10-20 parts of ginseng, 10-20 parts of platycodon grandiflorum, 8-16 parts of mulberry leaves, 8-16 parts of chrysanthemum, 8-16 parts of reed rhizome, 5-10 parts of lophatherum gracile, 5-10 parts of gardenia, 5-10 parts of dandelion, 5-10 parts of dried ginger, 7-14 parts of coix seeds, 6-12 parts of orange peels, 4-8 parts of liquorice and 4-8 parts of hawthorn.
As a preferred scheme, the method for detecting the fingerprint of the instant momordica grosvenori solid beverage comprises the following steps: 15 parts of momordica grosvenori, 10 parts of ginseng, 10 parts of platycodon grandiflorum, 8 parts of mulberry leaf, 8 parts of chrysanthemum, 8 parts of reed rhizome, 5 parts of lophatherum gracile, 5 parts of gardenia, 5 parts of dandelion, 5 parts of dried ginger, 7 parts of coix seed, 6 parts of orange peel, 4 parts of liquorice and 4 parts of hawthorn.
Optimizing fingerprint detection conditions:
1. in the aspect of preparation optimization of sample solution
According to the invention, through experimental comparison of different extraction methods (ultrasonic extraction, reflux extraction, percolation extraction and the like) and different extraction solvents (methanol, water, 50% ethanol water solution, 70% ethanol, 95% ethanol and absolute ethanol), the results show that the spectrogram difference component obtained by ultrasonic extraction is relatively comprehensive, the extraction efficiency is high, the separation degree is good, so that the ultrasonic extraction method is adopted; in the investigation of the extraction solvent, the chromatogram of the methanol extract is found to have the most information content and the highest component content, so that methanol is selected for extraction.
2. In the aspect of optimizing chromatographic conditions
According to the invention, a diode array detector is adopted to inspect the detection wavelength, chromatograms at positions of 205nm, 225nm, 245nm and 280nm are extracted, and when the detection wavelength is 280nm, the information content contained in the chromatograms is most comprehensive and the base line is stable, so that 280nm is selected as the detection wavelength;
the invention screens the flow rate (1mL/min, 0.9mL/min, 0.8mL/min and 0.6mL/min), because the components in the instant momordica grosvenori solid beverage mostly contain saponin, flavone and terpenoid isomerides and other components with very similar polarity, the components can not be separated at high flow rate, the separation effect is better at low flow rate, and finally the substances with similar polarity are separated under the gradient condition of the flow rate of 0.6 mL/min.
The invention compares the elution effects of 5 different elution systems of methanol-water, acetonitrile-0.1% formic acid, acetonitrile-0.1% phosphoric acid water and acetonitrile-0.2% phosphoric acid water under different gradients. As a result, it was found that acetonitrile and 0.2% phosphoric acid water were finally selected as the mobile phase because the separation effect of each component in the instant Siraitia grosvenorii solid beverage was better when acetonitrile and 0.2% phosphoric acid water were used as the mobile phase.
After the optimal fluidity composition is determined, the optimal gradient elution program is screened through a large number of experiments, and the experiment shows that when acetonitrile with the volume of 5% is adopted for 0-15 min; 5-21% of acetonitrile in 15-65 min; the volume of acetonitrile is 21% in 65-85 min; the volume of acetonitrile is 21-22% in 85-95 min; the volume of acetonitrile is 22-40% in 95-110 min; good separation degree of each spectrum peak in the fingerprint spectrum can be realized.
Has the advantages that:
1. according to the structural property characteristics of active ingredients contained in the instant momordica grosvenori solid beverage, the optimal mobile phase composition is screened out through a large number of experiments, and analysis conditions such as gradient elution procedures, flow rate, detection wavelength, chromatographic column, column temperature and the like are verified through a plurality of experiments.
2. The fingerprint detection method for the instant momordica grosvenori solid beverage provided by the invention can comprehensively, objectively and accurately detect and evaluate the quality of the instant momordica grosvenori solid beverage, and has important significance for ensuring the curative effect of the instant momordica grosvenori solid beverage.
3. The fingerprint of the instant momordica grosvenori solid drink established by the method can effectively represent the quality of the instant momordica grosvenori solid drink, objectively reflect the front and back sequence and the mutual relation of characteristic peaks of various fingerprints, pay attention to the overall facial features, avoid the one-sidedness of judging the quality of the instant momordica grosvenori solid drink due to the determination of individual chemical components, and reduce the possibility of manual processing for reaching the quality standard.
4. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage provided by the invention has the advantages of simplicity, convenience, good stability, high precision, good reproducibility and the like.
Drawings
FIG. 1 is a chromatogram of a mixed control according to the present invention.
FIG. 2 is a comparison fingerprint of the instant Momordica grosvenori Swingle solid beverage sample of the present invention.
FIG. 3 shows the fingerprints of 8 batches of samples of the instant Momordica grosvenori Swingle solid beverage of the present invention.
Detailed Description
Embodiments of the present invention will be described in detail with reference to examples, in which specific conditions are not specified, according to conventional conditions or conditions recommended by manufacturers. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
The following examples used the following instruments and reagents:
experimental equipment
1.1 instruments
A high-performance liquid chromatography system with full-wave-band scanning (200- & 800nm) of Shimadzu corporation in Japan comprises a full-automatic online degassing system, a full-automatic sample introduction system promience SIL-20A, a diode array detector SPD-M20A and an automatic temperature control column temperature box CTO-20A, a KQ3200DB type numerical control ultrasonic cleaner (Kunshan ultrasonic instruments Co., Ltd.), and a BP121S electronic analytical balance (SARTORIUS).
1.2 drugs and reagents
The instant fructus Siraitiae Grosvenorii solid beverage sample is from Jiangsu Haohai pharmaceutical industry Co., Ltd; chlorogenic acid reference (batch No. 110753-201314) was purchased from China food and drug Biometrics institute; the hesperidin control (batch C-024-130701) was purchased from Douglas Biotech, Inc.; methanol (analytically pure); phosphoric acid (analytically pure); acetonitrile (chromatographically pure); water (ultrapure water).
Embodiment 1 a method for detecting a fingerprint of instant momordica grosvenori solid beverage, comprising the following steps:
taking 25g (3.25 g of momordica grosvenori, 2.5g of ginseng, 2.5 parts of platycodon grandiflorum, 2 parts of mulberry leaf, 2 parts of chrysanthemum, 2 parts of reed rhizome, 1.25 parts of lophatherum gracile, 1.25 parts of gardenia, 1.25 parts of dandelion, 1.25 parts of dried ginger, 1.75 parts of coix seed, 1.5 parts of orange peel, 1 part of liquorice and 1 part of hawthorn) of an instant momordica grosvenori solid beverage clear paste sample of 8 batches, then placing the sample in a 500mL round-bottomed bottle, adding 300mL of methanol solution, ultrasonically extracting for 30min, centrifuging 12000 for 5min, taking supernatant, and filtering the supernatant through a 0.45 mu m microfiltration membrane to obtain a test solution;
precisely weighing chlorogenic acid and hesperidin, placing in a volumetric flask, adding methanol to desired volume to scale, shaking, and making into mixed reference solution containing 0.1019mg/mL chlorogenic acid and 0.1101mg/mL hesperidin.
| Procedure | time (min) | Acetonitrile volume (%) |
| 1 | 0 | 5 |
| 2 | 15 | 5 |
| 3 | 65 | 21 |
| 4 | 85 | 21 |
| 5 | 95 | 22 |
| 6 | 110 | 40 |
Step 4, exporting the fingerprints of the 8 batches of instant momordica grosvenori solid beverage test sample solutions obtained in the step 3, and importing the fingerprints into a traditional Chinese medicine chromatography fingerprint similarity evaluation system 2012A; selecting chromatographic peaks existing in chromatograms of 8 batches of instant siraitia grosvenorii solid beverages as common peaks; generating a reference fingerprint of 1 batch of instant siraitia grosvenorii solid beverage by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; as a result, 14 common peaks exist in 1 batch of crude instant fructus Siraitiae Grosvenorii solid beverage, the reference fingerprint spectrum is shown in FIG. 2, and the fingerprint spectrum of 8 batches of the test sample is shown in FIG. 3. The retention time of chlorogenic acid is 43.729min, and the retention time of hesperidin is 69.55 min.
And 5, comparing the fingerprint spectrum of the instant momordica grosvenori solid beverage obtained in the step 3 with the spectrum of the mixed standard substance (shown in figure 1), identifying the main components, and comparing the number 6 and 10 chromatographic peaks in the instant momordica grosvenori solid beverage as follows: chlorogenic acid (retention time 43.729min), and hesperidin (retention time 69.55 min).
Meanwhile, the invention uses the automatically generated reference HPLC fingerprint spectrum R to generate a common chromatographic peak mode, and the common chromatographic peaks of 8 batches of instant siraitia grosvenori solid beverage traditional Chinese medicines of Jiangsu Heishao pharmaceutical manufacturers obtained by analysis and calculation have relatively good similarity, which shows that the fingerprint spectrum established by the instant siraitia grosvenori solid beverage traditional Chinese medicines established by the method can well detect the quality of multiple batches of instant siraitia grosvenori solid beverages in Jiangsu Heishao pharmaceutical industries, and the results are shown in Table 1.
TABLE 1 similarity between batches of samples and consensus patterns
| S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | R | |
| S1 | 1.000 | 1.000 | 0.983 | 0.955 | 0.982 | 0.964 | 0.964 | 0.979 | 0.985 |
| S2 | 1.000 | 1.000 | 0.983 | 0.955 | 0.982 | 0.964 | 0.964 | 0.979 | 0.985 |
| S3 | 0.983 | 0.983 | 1.000 | 0.976 | 0.997 | 0.980 | 0.976 | 0.992 | 0.995 |
| S4 | 0.955 | 0.955 | 0.976 | 1.000 | 0.976 | 0.991 | 0.960 | 0.983 | 0.987 |
| S5 | 0.982 | 0.982 | 0.997 | 0.976 | 1.000 | 0.981 | 0.978 | 0.992 | 0.995 |
| S6 | 0.964 | 0.964 | 0.980 | 0.991 | 0.981 | 1.000 | 0.965 | 0.987 | 0.992 |
| S7 | 0.964 | 0.964 | 0.976 | 0.960 | 0.978 | 0.965 | 1.000 | 0.970 | 0.982 |
| S8 | 0.979 | 0.979 | 0.992 | 0.983 | 0.992 | 0.987 | 0.970 | 1.000 | 0.996 |
| R | 0.985 | 0.985 | 0.995 | 0.987 | 0.995 | 0.992 | 0.982 | 0.996 | 1.000 |
Example 2 methodological study of fingerprint detection
1. Methodology investigation
1.1 precision investigation
Taking 25g of an instant momordica grosvenori solid beverage sample (the same medicinal ingredients and proportion as in example 1), preparing a sample solution according to the sample preparation method of example 1, carrying out continuous sample injection for 6 times, wherein the sample injection amount is 10 mu L each time, detecting according to the chromatographic conditions of example 1, determining an HPLC chromatogram, and inspecting 14 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 0.9%, the common peak area RSD is less than 1.6%, and the precision of the instrument is better.
1.2 stability Studies
Taking 25g of an instant momordica grosvenori solid beverage sample (the same medicinal taste composition and proportion as in example 1), preparing a sample solution according to the sample preparation method of example 1, injecting samples for 0, 2, 4, 8, 12 and 24 hours according to the chromatographic conditions in example 1, recording a chromatogram, and inspecting 14 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 1.2%, the common peak area RSD is less than 2.2%, and the sample solution has better stability within 24 hours.
1.3 repeatability test
Taking 6 parts of instant momordica grosvenori solid beverage sample, preparing a sample solution according to the sample preparation method in the example 1, respectively measuring, recording a chromatogram, and inspecting 14 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 2.1%, and the area RSD of the common fingerprint peaks is less than 2.5%, which indicates that the method has good repeatability.
The experimental results show that the fingerprint detection method for the instant momordica grosvenori solid beverage provided by the invention has the advantages of good stability, high precision and good repeatability, can comprehensively and objectively evaluate the quality of the instant momordica grosvenori solid beverage, and has important significance for ensuring the clinical curative effect.
The above embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and the scope of the present invention is defined by the claims. Various modifications and equivalents may be made by those skilled in the art within the spirit and scope of the present invention, and such modifications and equivalents should also be considered as falling within the scope of the present invention.
Claims (7)
1. A method for detecting fingerprint spectrum of instant momordica grosvenori solid beverage is characterized by comprising the following steps:
step 1, preparing a sample solution of instant momordica grosvenori solid beverage:
weighing different batches of instant fructus Siraitiae Grosvenorii solid beverage fluid extract samples, adding methanol solution, ultrasonic extracting, centrifuging the extractive solution, and filtering the supernatant with 0.45 μm microporous membrane to obtain sample solution;
step 2, preparation of mixed reference solution:
precisely weighing chlorogenic acid and hesperidin as reference substances, placing in a volumetric flask, adding methanol to desired volume to scale, shaking to obtain mixed reference substance solution;
step 3, precisely absorbing the test solution obtained in the step 1 and the mixed reference solution obtained in the step 2 respectively, injecting the solutions into a high performance liquid chromatograph, and recording a chromatogram;
step 4, exporting the fingerprint of the instant momordica grosvenori solid beverage test solution obtained in the step 3, and importing the fingerprint into a traditional Chinese medicine chromatography fingerprint similarity evaluation system 2004A; selecting chromatographic peaks existing in chromatograms of different batches of instant siraitia grosvenorii solid beverages as common peaks; generating a comparison fingerprint of the instant momordica grosvenori solid beverage by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; marking chemical components of peaks in the comparison fingerprint spectrum according to the retention time of the mixed reference solution chromatogram;
and 5, comparing the fingerprint spectrum of the instant momordica grosvenori solid beverage obtained in the step 3 with the spectrum of the mixed standard substance, and identifying that the peak 6 in the instant momordica grosvenori solid beverage is chlorogenic acid and the peak 10 in the instant momordica grosvenori solid beverage is hesperidin.
2. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to claim 1, wherein the preparation method of the sample solution of the instant momordica grosvenori solid beverage in the step 1 comprises the following steps: taking 5g of 8 batches of instant siraitia grosvenorii solid beverage clear paste samples, placing the samples in a round-bottom flask, carrying out ultrasonic extraction, centrifuging an extracting solution, and filtering supernate with a 0.45 mu m microporous filter membrane to obtain a sample solution.
3. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to claim 1, wherein the step 2 is to prepare a mixed reference solution: precisely weighing chlorogenic acid and hesperidin, placing in a volumetric flask, adding methanol to desired volume to scale, shaking, and making into mixed reference solution containing 0.1019mg/mL chlorogenic acid and 0.1101mg/mL hesperidin.
4. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to claim 1, wherein in the step 3, the liquid chromatography conditions are as follows: a chromatographic column: YMC-Pack ODS-A, mobile phase: acetonitrile and 0.2% phosphoric acid water, gradient elution, diode array detector, detection wavelength: 210nm, column temperature 30 ℃, flow rate 0.6mL/min, sample injection volume: 10 μ L, gradient elution procedure as follows:
5. the method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to claim 1, wherein the fingerprint contains 14 peaks, and the retention time of the 14 peaks is 20.329min for peak 1, 25.649min for peak 2, 28.81min for peak 3, 32.186min for peak 4, 38.657min for peak 5, 43.729min for peak 6, 45.644min for peak 7, 47.377min for peak 8, 49.905min for peak 9, 69.55min for peak 10, 73.524min for peak 11, 77.167min for peak 12, 80.223min for peak 13 and 84.691min for peak 14, wherein the peak 6 is chlorogenic acid and the peak 10 is hesperidin.
6. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to any one of claims 1 to 5, wherein the instant momordica grosvenori solid beverage comprises the following medicinal ingredients in parts by weight: 15-30 parts of momordica grosvenori, 10-20 parts of ginseng, 10-20 parts of platycodon grandiflorum, 8-16 parts of mulberry leaves, 8-16 parts of chrysanthemum, 8-16 parts of reed rhizome, 5-10 parts of lophatherum gracile, 5-10 parts of gardenia, 5-10 parts of dandelion, 5-10 parts of dried ginger, 7-14 parts of coix seeds, 6-12 parts of orange peels, 4-8 parts of liquorice and 4-8 parts of hawthorn.
7. The method for detecting the fingerprint of the instant momordica grosvenori solid beverage according to claim 6, wherein the instant momordica grosvenori solid beverage comprises the following medicinal ingredients in parts by weight: 15 parts of momordica grosvenori, 10 parts of ginseng, 10 parts of platycodon grandiflorum, 8 parts of mulberry leaf, 8 parts of chrysanthemum, 8 parts of reed rhizome, 5 parts of lophatherum gracile, 5 parts of gardenia, 5 parts of dandelion, 5 parts of dried ginger, 7 parts of coix seed, 6 parts of orange peel, 4 parts of liquorice and 4 parts of hawthorn.
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