CN114062553A - Method for measuring HPLC fingerprint of child six-ingredient appetite increasing ointment - Google Patents
Method for measuring HPLC fingerprint of child six-ingredient appetite increasing ointment Download PDFInfo
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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
- G01N30/02—Column chromatography
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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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- G01N30/06—Preparation
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- 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
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- 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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- 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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Abstract
The invention belongs to the field of pharmaceutical analysis of a six-ingredient appetite promoting paste for children, and particularly relates to a method for measuring an HPLC (high performance liquid chromatography) fingerprint spectrum of the six-ingredient appetite promoting paste for children, which is characterized in that a preparation method of a test solution determined through the investigation of symmetric sample amount, an extraction solvent, an extraction mode and extraction time is as follows: heating, refluxing and extracting the child six-ingredient appetite-enhancing paste, filtering, and taking a subsequent filtrate to obtain a test solution; measuring by high performance liquid chromatography with gradient elution to obtain finger print of the children's six-ingredient appetite promoting paste, wherein the detection wavelength of the high performance liquid chromatography is 0-7 min:215 nm; 7-78 min, 330nm, and the flow rate is 0.8 mL/min. The fingerprint of the child six-ingredient appetite-enhancing paste is established by the high performance liquid chromatography, the fingerprint has 10 common peaks, the method is strong in specificity, high in precision, good in repeatability and high in stability, the purpose of effectively controlling the quality of the medicine is achieved, and the stability of the quality of the product is ensured.
Description
Technical Field
The invention belongs to the field of pharmaceutical analysis of a six-ingredient appetite promoting paste for children, and particularly relates to a method for measuring an HPLC (high performance liquid chromatography) fingerprint spectrum of the six-ingredient appetite promoting paste for children.
Background
Infantile anorexia refers to anorexia, or even refusal to eat. Anorexia patients often have the clinical manifestations of pale complexion, spontaneous perspiration, night sweat, nausea, vomiting, abdominal pain, abdominal distension, abnormal stool, emaciation, allotropic addiction and the like. The children patients are weak in constitution and low in immune function, and are susceptible to cold or oral ulcer. Severe cases may be manifested by "malnutritional" with symptoms of listlessness, thin neck, abdominal distention, and dry skin. The disease is delayed and not cured, and the growth, development and nutritional state of the children are influenced; in recent years, the number of diseases tends to increase in cities.
The theory of traditional Chinese medicine considers that: the spleen is the acquired root, the digestion and absorption functions are important for the abnormality of the whole human body, the good functions of the digestive system are kept, and the method is an important measure for ensuring the nutrition supply of the whole body and maintaining the growth, development and metabolism.
TCM believes that the disease is caused by incoordination between the spleen and stomach. The spleen and stomach are not harmonized with each other, so that the food cannot be eaten, and the essence cannot be distributed on four sides to nourish the whole body, so that the body is weak and thin, the face is sallow, the skin and the hair are hagky, the head and the neck are thin, the abdominal distention is full of the abdomen or the belly is sunken like a boat, and the patient feels restless and has acute anorexia. If the treatment is not timely performed, deficiency of qi and blood will be caused to affect the growth and development of the children patients. The short course of disease is usually an excess syndrome, the excess includes the stagnation of intestine and stomach, phlegm obstruction of middle energizer and worm accumulation, and it should be treated by differentiation and differentiation in clinic, mainly to regulate qi, promote digestion and remove stagnation; if the disease course is longer, it is usually indicated as deficiency syndrome, including dysfunction of spleen and stomach, deficiency of spleen and stomach qi and deficiency of stomach yin, or both deficiency and excess syndromes, it is mainly indicated for tonifying qi, strengthening spleen and tonifying deficiency, or both tonics and purgatives.
The six-ingredient appetite-enhancing paste for children comprises radix Ophiopogonis, rhizoma Polygonati Odorati, rhizoma Dioscoreae, fructus crataegi, radix asparagi, and Arecae semen, and adjuvants including Mel and sodium benzoate. The functional indications are as follows: nourishing yin, invigorating spleen, removing food stagnation, and lowering qi; it is used for treating infantile anorexia. Based on the pathogenesis of spleen deficiency and yin deficiency and milk food stagnation, the radix ophiopogonis and the polygonatum are used as monarch medicines in the formula to nourish the stomach and promote the production of body fluid; chinese yam is used for nourishing yin and tonifying spleen (Chinese famous traditional medicine Zhang Xin is … 'Chinese yam is used for nourishing stomach yin, gastric juice is sufficient, and people can take food by themselves' …); radix asparagi is added to strengthen the stomach's ability to promote the production of body fluid, strengthen the middle-jiao, promote the production of body fluid and nourish the fluid, and the six fu-organs are communicated as tonics; the charred hawthorn and the betel nut are used for removing stagnation, clearing hollow viscera and lowering stomach qi, so as to achieve the effect of tonifying and dredging. Tonify yin and dredge fu organs, so it has good therapeutic effect on anorexia, dry mouth, constipation and abdominal distention.
The fingerprint is a quantifiable and visual traditional Chinese medicine detection method, can be used for identifying authenticity and evaluating the uniformity and stability of the quality of traditional Chinese medicines, and the HPLC fingerprint technology is widely applied to the field of quality control of traditional Chinese medicines, however, at present, the quality control aspect of the child six-ingredient appetite-enhancing ointment is not reported and recorded by the HPLC fingerprint method.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the method overcomes the defects of the prior art, provides a method for measuring the HPLC fingerprint of the children's six-ingredient appetite increasing paste, establishes the standard fingerprint of the children's six-ingredient appetite increasing paste, and can monitor the quality of the children's six-ingredient appetite increasing paste more comprehensively.
In order to realize the purpose, the invention is realized by adopting the following technical scheme:
according to the invention, a preparation method of a test sample solution is determined through the investigation of the amount of a symmetrical sample, an extraction solvent, an extraction mode and extraction time, a fingerprint is established by adopting a high performance liquid chromatography, and the test sample fingerprint is evaluated by adopting a traditional Chinese medicine chromatography fingerprint similarity evaluation system (2012A edition) published by the State pharmacopoeia Committee.
The HPLC fingerprint spectrum determination method of the children's six-ingredient appetite increasing paste comprises the steps of heating, refluxing, extracting and filtering the children's six-ingredient appetite increasing paste, and taking a subsequent filtrate to obtain a test solution; measuring by high performance liquid chromatography with gradient elution to obtain finger print of the children's six-ingredient appetite promoting paste, wherein the detection wavelength of the high performance liquid chromatography is 0-7 min:215 nm; 7-78 min, 330nm, and the flow rate is 0.8 mL/min. The determination was carried out as follows:
(1) chromatographic conditions and system applicability: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature: the number of theoretical plates is not less than 3000 calculated by chlorogenic acid peak at 30 ℃;
(2) preparation of a reference solution: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL;
(3) preparing a reference medicinal material solution:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution;
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference medicinal material solution;
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue with 25mL of 30% methanol to obtain reference material solution;
fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
(4) preparing a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, and collecting the filtrate;
(5) preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
(6) and (3) determination: precisely sucking 10 μ L of each of the reference solution, the reference medicinal material solution, the negative solution and the sample solution, injecting into high performance liquid chromatograph, and measuring.
Preferably, the HPLC fingerprint of the children's six-ingredient appetite-enhancing paste is determined by heating, refluxing, extracting, filtering, taking a subsequent filtrate, decolorizing with a novel material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, concentrating and drying to obtain a test solution, and determining by high performance liquid chromatography of gradient elution to obtain the fingerprint of the children's six-ingredient appetite-enhancing paste, wherein the detection wavelength of the high performance liquid chromatography is 0-7 min:215 nm; 7-78 min, 330nm, and the flow rate is 0.8 mL/min. The determination was carried out as follows:
(1) chromatographic conditions and system applicability: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature: the number of theoretical plates is not less than 3000 calculated by chlorogenic acid peak at 30 ℃;
(2) preparation of a reference solution: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL;
(3) preparing a reference medicinal material solution:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution;
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference medicinal material solution;
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue with 25mL of 30% methanol to obtain reference material solution;
fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
(4) preparing a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
(5) preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
(6) and (3) determination: precisely sucking 10 μ L of each of the reference solution, the reference medicinal material solution, the negative solution and the sample solution, injecting into high performance liquid chromatograph, and measuring.
The specific conditions of the gradient elution are as follows: 0-25 min: a and B are 5-20: 95-80; 25-45 min: a and B are 20-25: 80-75; 45-58 min: a and B are 25-40: 75-60; 58-65 min: a and B are 40-45: 60-55; 65-68 min: a and B are 45-5: 55-95; 68-78 min: and A and B are 5: 95.
There are 10 common peaks in the fingerprint, and the serial number is 1 ~ 10 in proper order: peak 9 (chlorogenic acid) was taken as the S peak, and the peak numbers (relative retention times) of the common peaks were recorded as: 1(0.19), 2(0.22), 3(0.26), 4(0.34), 5(0.43), 6(0.67), 7(0.79), 8(0.87), 9(1.00), 10 (1.07); peak 5 is 5-hydroxymethylfurfural and peak 9(S) is chlorogenic acid.
The main contents of the relevant experimental research and investigation carried out by the invention are as follows:
an instrument and equipment
TABLE 1 instruments and apparatus
Medicine and reagent
TABLE 2 drugs and reagents
Third, method and results
1. Selection of chromatographic conditions
1.1 selection of different wavelengths
Repeated tests on the wavelength of the six medicinal materials in the child six-ingredient appetite-enhancing paste show that the wavelength is 215nm in 0-7 min; the maximum absorption wavelength is 330nm at 7-78 min, so the detection wavelength of the six-ingredient appetite-enhancing paste for children is determined to be 0-7 min:215 nm; at 330nm in the range of 7-78 min, and the chromatogram is shown in figure 1.
1.2 column temperature selection
In order to find the optimum column temperature, experiments were carried out using different column temperatures, the chromatogram is shown in FIG. 2, and the results are shown in Table 3.
TABLE 3 investigation of different column temperatures
And (4) conclusion: as a result, it was found that 25 ℃ or 30 ℃ can be selected.
1.3 flow Rate selection
To find the optimum flow rate, the applicant performed experiments using different flow rates, the chromatogram being shown in FIG. 3 and the results in Table 4.
Table 4 different flow rate investigation
And (4) conclusion: from the results, it was found that the separation degree of each chromatographic peak was good, the peak shape was good and the effect was best at a flow rate of 0.8 mL/min.
1.4 selection of different chromatography columns
In order to find the best column, the applicant performed tests using different columns, the results of which are shown in table 5.
Table 5 different column investigation
And (4) conclusion: from the results, it was found that different columns had a small influence on the degree of separation, but Welchrom C18 column showed a good peak shape at peak 1 and a relatively good degree of separation, and Welchrom C18 column was selected.
2. Preparation of control solutions
2.1 taking a proper amount of chlorogenic acid reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 140 μ g of chlorogenic acid per 1 mL.
2.2 taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 140 micrograms per 1 mL.
3. Preparation of test solution
3.1 investigation of the different sample weights
The method comprises the following steps: taking 1g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the second method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the third method comprises the following steps: taking 5g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol. The results are shown in Table 6 and the chromatograms are shown in FIG. 4.
TABLE 6 investigation of different sample weights
And (4) conclusion: through investigation of different sample weighing amounts, the second method has high extraction rate and good separation degree, and is more favorable for ensuring the accuracy and stability of the detection result.
3.2 investigation of different extraction solvents
The method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the second method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, accurately weighing, placing in a 150mL conical flask with a plug, accurately adding 50mL of methanol, weighing, heating and refluxing for 90min, cooling, and supplementing with 30% methanol to reduce weight loss; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the third method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of water, weighing, heating and refluxing for 90min, cooling, and supplementing with 30% methanol to reduce weight loss; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol. The results are shown in Table 7 and the chromatograms are shown in FIG. 5.
TABLE 7 investigation of different extraction solvents
And (4) conclusion: through investigation of different extraction solvents, the method I has high extraction rate and good separation degree, and is more favorable for ensuring the accuracy and stability of detection results.
3.3 examination of different extraction modes
The method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the second method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, ultrasonically extracting for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol. The results are shown in Table 8 and the chromatogram is shown in FIG. 6.
TABLE 8 examination of different extraction modes
And (4) conclusion: through investigation of different extraction modes, the method I has high extraction rate and good separation degree, and is more favorable for ensuring the accuracy and stability of detection results.
3.4 investigation of different extraction times
The method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 60min, and supplementing with 30% methanol to reduce weight loss; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the second method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, and supplementing with 30% methanol to reduce the weight loss; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
the third method comprises the following steps: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 120min, and supplementing with 30% methanol to reduce the weight loss; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol. The results are shown in Table 9 and the chromatograms are shown in FIG. 7.
TABLE 9 different extraction time survey
And (4) conclusion: through investigation of different extraction times, the extraction rate is higher and the separation degree is good in the second method and the third method, but the extraction time is longer in the third method, so that the second method is selected.
4. Stability of
Preparing a sample solution, precisely sucking 10 μ L of each sample at room temperature for 0, 2, 6, 10, and 22h, respectively, sampling for 6 times, calculating RSD, and showing the results in tables 10 and 11, and the chromatogram in FIG. 8.
TABLE 10 stability versus Retention time test results
TABLE 11 results of stability relative peak area test
And (4) conclusion: as is clear from the test results, the test solution was stable at room temperature for 22 hours.
5. Repeatability of
6 test solutions were prepared in parallel, 10. mu.L of each sample was injected, and RSD was calculated, and the results are shown in tables 12 and 13, and the chromatogram is shown in FIG. 9.
TABLE 12 repeatability versus retention time test results
TABLE 13 repeatability relative peak area test results
And (4) conclusion: from the test results, the method was found to have good reproducibility.
6. Precision degree
Preparing a sample solution, continuously injecting sample 6 needles in the same high performance liquid chromatograph, with sample amount of 10 μ L, calculating RSD, and obtaining results shown in tables 14 and 15 and chromatogram shown in figure 10.
TABLE 14 precision vs. Retention time test results
TABLE 15 results of relative peak area test for precision
And (4) conclusion: as is clear from the test results, the precision was good.
7. Investigation of different batches of test articles
Taking three batches of child six-ingredient appetite-enhancing paste (batch numbers: 210101, 210601 and 210603) 3g each, precisely weighing, placing in a conical flask with a plug of 150mL, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, and supplementing with 30% methanol to reduce weight loss; filtering, collecting filtrate, decolorizing with JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water and 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, dissolving with 30% methanol, and performing sample injection determination, wherein the result is shown in Table 16 and the chromatogram is shown in FIG. 11.
TABLE 16 results of relative peak area test for precision
And (4) conclusion: the test results show that the common peaks of different batches of the test sample are the same, and the reproducibility is good.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
Compared with the prior art, the invention has the following beneficial effects:
the fingerprint of the child six-ingredient appetite-enhancing paste and the standard fingerprint thereof are established for the first time by utilizing the HPLC technology, the method has strong specificity, high precision, good repeatability and high stability, the aim of effectively controlling the quality of the medicine is fulfilled, and the stability, safety and effectiveness of the quality of the product are ensured.
Drawings
FIG. 1 shows HPLC finger-prints of six-ingredient appetite-enhancing paste for children with different wavelengths (S1-S5 are chromatogram of test sample, chromatogram of wavelength 330nm, chromatogram of wavelength 280nm, chromatogram of wavelength 215nm and chromatogram of wavelength 203nm, respectively, from bottom to top);
FIG. 2 shows different column temperature HPLC finger prints of the children's six ingredient appetite-enhancing paste (30 ℃, 35 ℃ and 25 ℃ chromatograms from bottom to top);
FIG. 3 shows HPLC fingerprints (from bottom to top, 0.8mL/min chromatogram, 0.9mL/min chromatogram, and 0.7mL/min chromatogram) of different flow rates of the children's six-ingredient appetite-enhancing paste;
FIG. 4 shows HPLC finger prints of the six-ingredient appetite-enhancing paste for children in different sample weights (from bottom to top, 3g chromatogram, 5g chromatogram, 1g chromatogram);
FIG. 5 shows HPLC finger prints of different extraction solvents of the children's six-ingredient appetite-enhancing ointment (from bottom to top, 30% methanol extraction chromatogram, and water extraction chromatogram, respectively);
FIG. 6 shows HPLC finger prints of the six-ingredient appetite-enhancing paste for children in different extraction modes (from bottom to top, heating reflux extraction chromatogram and ultrasonic extraction chromatogram);
FIG. 7 is HPLC finger prints of the children's six-ingredient appetite-enhancing paste at different extraction times (from bottom to top, respectively, a chromatogram of 90min, a chromatogram of 60min, and a chromatogram of 120 min);
FIG. 8 shows a stability HPLC fingerprint of the pediatric six-ingredient appetite-enhancing paste (S1-S6 are respectively a chromatogram of 0h, a chromatogram of 2h, a chromatogram of 4h, a chromatogram of 8h, a chromatogram of 12h, and a chromatogram of 24h from bottom to top);
fig. 9 is a repetitive HPLC fingerprint of the pediatric six-ingredient appetite increasing paste (S1-S6 are, from bottom to top, a repetitive 1 chromatogram, a repetitive 2 chromatogram, a repetitive 3 chromatogram, a repetitive 4 chromatogram, a repetitive 5 chromatogram, and a repetitive 6 chromatogram, respectively);
FIG. 10 shows a precision HPLC fingerprint of the Xiaoer Liuwei Zengshi ointment (S1-S6 are respectively a precision 1 chromatogram, a precision 2 chromatogram, a precision 3 chromatogram, a precision 4 chromatogram, a precision 5 chromatogram, and a precision 6 chromatogram from bottom to top);
FIG. 11 is a HPLC fingerprint of different batches of the pediatric six-ingredient appetite-enhancing paste (from bottom to top, 210603, 210101 and 210601 chromatograms respectively);
FIG. 12 HPLC fingerprint of the infantile Liuwei Zengying paste;
FIG. 13 HPLC finger print of LIUWEIZENGSHI paste for children (S1-S3 from bottom to top are respectively a test solution, a chlorogenic acid reference solution, and a 5-hydroxymethylfurfural reference solution);
FIG. 14 shows HPLC finger prints of control drugs of LIUWEIZENGSHI ointment for children (from bottom to top, respectively, test solution, Arecae semen control solution, radix asparagi control solution, fructus crataegi control solution, rhizoma Polygonati Odorati control solution, and radix Ophiopogonis control solution);
FIG. 15 HPLC finger print of the specific HPLC fingerprint of the children' S Liuwei Zengshi ointment (S1-S3 are respectively the sample solution, honey and sodium benzoate from bottom to top).
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1
1. Chromatographic conditions
Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature 30 ℃, sample injection amount of 10 mu L, and theoretical plate number not less than 3000 according to chlorogenic acid peak. The method for measuring the HPLC fingerprint of the child six-ingredient appetite-enhancing paste comprises the following specific conditions of gradient elution: 0-25 min: a and B are 5-20: 95-80; 25-45 min: a and B are 20-25: 80-75; 45-58 min: a and B are 25-40: 75-60; 58-65 min: a and B are 40-45: 60-55; 65-68 min: a and B are 45-5: 55-95; 68-78 min: and A and B are 5: 95.
2. Preparation of reference substance, reference medicinal material, test sample and negative solution
Preparation of control solutions: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL.
Preparing reference medicinal materials:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution.
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a reference medicinal material solution.
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating the filtrate to dryness, and dissolving the residue in 25mL of 30% methanol to obtain a reference material solution.
Fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution.
Fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference material solution.
Preparation of a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol.
Preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
3. assay method
Respectively taking the reference substance, the reference medicinal material, the test sample and the negative solution, determining according to chromatographic conditions, injecting 10 mu L of sample, and recording chromatogram, wherein the results are shown in figures 12-15. According to results, the negative solution of the HPLC fingerprint of the child six-ingredient appetite-enhancing paste obtained under the conditions has no interference on a test sample, the preparation method of the test sample solution is simple, the number of peaks is large, and the separation degree of main characteristic peaks is good.
Identifying main chromatographic peaks by using a reference substance and a reference medicinal material, and judging that the peak 1 is a specific peak of the areca, the peak 3 is a specific peak of the areca, the peak 4 is a common peak of all the reference medicinal materials, the peak 5 is 5-hydroxymethylfurfural, the peak 6 is a common peak of the areca, the asparagus and the hawthorn, the peak 9 is chlorogenic acid and the peak 10 is a specific peak of the asparagus.
Example 2
1. Chromatographic conditions
Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature 30 ℃, sample injection amount of 10 mu L, and theoretical plate number not less than 3000 according to chlorogenic acid peak. The method for measuring the HPLC fingerprint of the child six-ingredient appetite-enhancing paste comprises the following specific conditions of gradient elution: 0-25 min: a and B are 5-20: 95-80; 25-45 min: a and B are 20-25: 80-75; 45-58 min: a and B are 25-40: 75-60; 58-65 min: a and B are 40-45: 60-55; 65-68 min: a and B are 45-5: 55-95; 68-78 min: and A and B are 5: 95.
2. Preparation of reference substance, reference medicinal material, test sample and negative solution
Preparation of control solutions: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL.
Preparing reference medicinal materials:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution.
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a reference medicinal material solution.
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating the filtrate to dryness, and dissolving the residue in 25mL of 30% methanol to obtain a reference material solution.
Fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution.
Fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference material solution.
Preparation of a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, and collecting the filtrate.
Preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
3. assay method
The reference substance, the reference medicinal materials, the test sample and the negative solution are respectively taken, the sample introduction is carried out by 10 mu L according to the chromatographic condition, the chromatogram is recorded, the result shows that the negative solution of the HPLC fingerprint of the child six-ingredient appetite promoting ointment obtained under the condition has no interference on the test sample, the preparation method of the test sample solution is simple, the number of peaks is large, and the separation degree of main characteristic peaks is good.
Identifying main chromatographic peaks by using a reference substance and a reference medicinal material, and judging that the peak 1 is a specific peak of the areca, the peak 3 is a specific peak of the areca, the peak 4 is a common peak of all the reference medicinal materials, the peak 5 is 5-hydroxymethylfurfural, the peak 6 is a common peak of the areca, the asparagus and the hawthorn, the peak 9 is chlorogenic acid and the peak 10 is a specific peak of the asparagus.
Example 3
Precisely sucking the test sample solution of the pediatric six-ingredient appetite-enhancing paste in example 1, respectively standing for 0h, 2h, 6h, 10h and 22h, measuring according to the chromatographic conditions in example 1, analyzing the relative retention time and the relative peak area of the characteristic peak by taking a chlorogenic acid peak as a reference peak (S peak), and calculating RSD.
Example 4
Precisely absorbing the test sample solution of the pediatric six-ingredient appetite-enhancing paste in example 1, measuring according to the chromatographic conditions in example 1, continuously injecting samples for 6 times, taking a chlorogenic acid peak as a reference peak (S peak), analyzing the relative retention time and the relative peak area of a characteristic peak, and calculating RSD.
Example 5
According to the preparation method of the test solution of the six-ingredient appetite-enhancing paste for children in example 1, 6 samples are prepared and injected respectively, the relative retention time and the relative peak area of the characteristic peak are analyzed by taking the chlorogenic acid peak as a reference peak (S peak), and the RSD is calculated.
Example 6
According to the preparation method of the test solution of the six-ingredient appetite-enhancing paste for children in the embodiment 1, sample injection is respectively carried out at the column temperatures of 25 ℃, 30 ℃ and 35 ℃, so that the peak shape and the separation degree of the test sample injected by the method with the column temperature of 25 ℃ and 30 ℃ are better, and the peak shape of the test sample injected by the method with the column temperature of 35 ℃ is poorer; therefore, the column temperature can be selected to be 25 ℃ and 30 ℃, and the chromatogram is shown in figure 2.
Example 7
According to the preparation method of the test solution of the child six-ingredient appetite-enhancing paste in the embodiment 1, the samples are respectively injected at the flow rates of 0.7ml/min, 0.8ml/min and 0.9ml/min, and the peak shape of the test sample injected at the flow rate of 0.7ml/min is poor; the separation degree of the sample injected at the flow rate of 0.9ml/min is poor; the sample with the flow rate of 0.8ml/min is injected with good peak shape and separation degree, so the flow rate of 0.8ml/min is selected for injection, and the chromatogram is shown in figure 3.
Of course, the foregoing is only a preferred embodiment of the invention and should not be taken as limiting the scope of the embodiments of the invention. The present invention is not limited to the above examples, and equivalent changes and modifications made by those skilled in the art within the spirit and scope of the present invention should be construed as being included in the scope of the present invention.
Claims (6)
1. A method for measuring HPLC fingerprint of child six-ingredient appetite-enhancing paste is characterized by comprising the following steps: heating, refluxing and extracting the child six-ingredient appetite-enhancing paste, filtering, and taking a subsequent filtrate to obtain a test solution; measuring by high performance liquid chromatography with gradient elution to obtain finger print of the children's six-ingredient appetite promoting paste, wherein the detection wavelength of the high performance liquid chromatography is 0-7 min:215 nm; 7-78 min, 330nm, and the flow rate is 0.8 mL/min.
2. The method for determining the HPLC fingerprint of the Liuwei Zengying paste for children according to claim 1, which is characterized in that: heating reflux extraction of the six-ingredient appetite-enhancing paste for the children, filtering, taking a subsequent filtrate, decoloring by a novel material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing by using IXD-100 macroporous adsorption resin, eluting by using water, eluting by using 70% ethanol, collecting 70% ethanol eluent, concentrating and drying to obtain a test solution, and measuring by using a high performance liquid chromatography of gradient elution to obtain a fingerprint of the six-ingredient appetite-enhancing paste for the children, wherein the detection wavelength of the high performance liquid chromatography is 0-7 min:215 nm; 7-78 min, 330nm, and the flow rate is 0.8 mL/min.
3. The method for determining the HPLC fingerprint of the Liuwei Zengying paste for children according to claim 1, which is characterized in that: the determination was carried out as follows:
(1) chromatographic conditions and system applicability: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature: the number of theoretical plates is not less than 3000 calculated by chlorogenic acid peak at 30 ℃;
(2) preparation of a reference solution: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL;
(3) preparing a reference medicinal material solution:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution;
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference medicinal material solution;
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue with 25mL of 30% methanol to obtain reference material solution;
fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
(4) preparing a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, and collecting the filtrate;
(5) preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
(6) and (3) determination: precisely sucking 10 μ L of each of the reference solution, the reference medicinal material solution, the negative solution and the sample solution, injecting into high performance liquid chromatograph, and measuring.
4. The method for determining the HPLC fingerprint of the Liuwei Zengying paste for children according to claim 2, which is characterized in that: the determination was carried out as follows:
(1) chromatographic conditions and system applicability: octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as a mobile phase A and 0.1% phosphoric acid aqueous solution as a mobile phase B, carrying out gradient elution, and detecting the wavelength by ultraviolet: 215nm for 0-7 min; 330nm for 7-78 min, flow rate: 0.8mL/min, column temperature: the number of theoretical plates is not less than 3000 calculated by chlorogenic acid peak at 30 ℃;
(2) preparation of a reference solution: precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 30% methanol to obtain solution containing 140 μ g per 1 mL; taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 30% methanol to prepare a solution containing 100 micrograms per 1 mL;
(3) preparing a reference medicinal material solution:
firstly, hawthorn: weighing 2g of hawthorn control medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue in 25mL of 30% methanol to prepare a control medicinal material solution;
② betel nut: weighing 2g of areca catechu reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare a reference medicinal material solution;
③ asparagus: weighing 2g of radix asparagi reference material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residue with 25mL of 30% methanol to obtain reference material solution;
fourthly, radix ophiopogonis: weighing 2g of radix ophiopogonis reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
fifthly, fragrant solomonseal rhizome: weighing 2g of polygonatum odoratum reference medicinal material, adding 100mL of water, decocting and concentrating to about 20mL, filtering, evaporating filtrate to dryness, and dissolving residues in 25mL of 30% methanol to prepare reference medicinal material solution;
(4) preparing a test solution: taking 3g of the child six-ingredient appetite-enhancing paste, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss with 30% methanol; filtering, collecting the filtrate, decolorizing with new material JHE8 hydrated magnesium aluminum silicate chromatographic column, adsorbing with IXD-100 macroporous adsorbent resin, eluting with water, eluting with 70% ethanol, collecting 70% ethanol eluate, recovering solvent, concentrating, drying, and dissolving with 30% methanol to obtain the final product;
(5) preparation of negative solution:
firstly, taking 0.6g of honey, precisely weighing, placing in a 150mL conical flask with a plug, precisely adding 50mL of 30% methanol, weighing, heating and refluxing for 90min, cooling, and supplementing the weight loss by using 30% methanol; filtering, and collecting the filtrate;
secondly, 0.09g of sodium benzoate is precisely weighed, is placed into a 150mL conical flask with a plug, 50mL of 30 percent methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 90min, is cooled, and is complemented with 30 percent methanol to reduce the weight loss; filtering, and collecting the filtrate;
(6) and (3) determination: precisely sucking 10 μ L of each of the reference solution, the reference medicinal material solution, the negative solution and the sample solution, injecting into high performance liquid chromatograph, and measuring.
5. The method for determining the HPLC fingerprint of the child Liuwei Zengying paste according to claim 1 or 2, wherein the HPLC fingerprint of the child Liuwei Zengying paste is as follows: the specific conditions of the gradient elution are as follows: 0-25 min: a and B are 5-20: 95-80; 25-45 min: a and B are 20-25: 80-75; 45-58 min: a and B are 25-40: 75-60; 58-65 min: a and B are 40-45: 60-55; 65-68 min: a and B are 45-5: 55-95; 68-78 min: and A and B are 5: 95.
6. The method for determining the HPLC fingerprint of the child Liuwei Zengying paste according to claim 1 or 2, wherein the HPLC fingerprint of the child Liuwei Zengying paste is as follows: there are 10 common peaks in the fingerprint, and the serial number is 1 ~ 10 in proper order: peak 9 (chlorogenic acid) was taken as the S peak, and the peak numbers (relative retention times) of the common peaks were recorded as: 1(0.19), 2(0.22), 3(0.26), 4(0.34), 5(0.43), 6(0.67), 7(0.79), 8(0.87), 9(1.00), 10 (1.07); peak 5 is 5-hydroxymethylfurfural and peak 9(S) is chlorogenic acid.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114609291A (en) * | 2022-03-22 | 2022-06-10 | 国药集团精方（安徽）药业股份有限公司 | Fingerprint detection method for compound Chinese medicinal decoction |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101444604A (en) * | 2009-01-06 | 2009-06-03 | 贵州本草堂药业有限公司 | Chinese medicine granular formulation to therapy children anorexia and preparation method thereof and detection method |
| CN102221590A (en) * | 2011-03-25 | 2011-10-19 | 湖南汉森制药股份有限公司 | Method for simultaneously determining multi-index ingredients of Simotang preparation and establishing fingerprint chromatogram thereof |
| CN102552751A (en) * | 2010-12-29 | 2012-07-11 | 江西济民可信集团有限公司 | Quality detection method for medicine (named as Er bao concentrated decoction) |
| 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 |
| CN104502518A (en) * | 2014-12-04 | 2015-04-08 | 吉林敖东延边药业股份有限公司 | Detection method of traditional Chinese medicine preparation for treating child anorexia |
| CN107153098A (en) * | 2016-03-03 | 2017-09-12 | 石家庄以岭药业股份有限公司 | A kind of assay method of the finger-print of Chinese medicine composition |
| WO2021123327A1 (en) * | 2019-12-19 | 2021-06-24 | Krka, D.D., Novo Mesto | Analytical method for iceland moss |
-
2021
- 2021-11-17 CN CN202111363640.5A patent/CN114062553B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101444604A (en) * | 2009-01-06 | 2009-06-03 | 贵州本草堂药业有限公司 | Chinese medicine granular formulation to therapy children anorexia and preparation method thereof and detection method |
| CN102552751A (en) * | 2010-12-29 | 2012-07-11 | 江西济民可信集团有限公司 | Quality detection method for medicine (named as Er bao concentrated decoction) |
| CN102221590A (en) * | 2011-03-25 | 2011-10-19 | 湖南汉森制药股份有限公司 | Method for simultaneously determining multi-index ingredients of Simotang preparation and establishing fingerprint chromatogram thereof |
| 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 |
| CN104502518A (en) * | 2014-12-04 | 2015-04-08 | 吉林敖东延边药业股份有限公司 | Detection method of traditional Chinese medicine preparation for treating child anorexia |
| CN107153098A (en) * | 2016-03-03 | 2017-09-12 | 石家庄以岭药业股份有限公司 | A kind of assay method of the finger-print of Chinese medicine composition |
| WO2021123327A1 (en) * | 2019-12-19 | 2021-06-24 | Krka, D.D., Novo Mesto | Analytical method for iceland moss |
Non-Patent Citations (1)
| Title |
|---|
| 江行本等: "HPLC法测定小儿增食片中大黄素和大黄酚的含量", 《河南中医学院学报》, vol. 20, no. 118, pages 32 - 33 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114609291A (en) * | 2022-03-22 | 2022-06-10 | 国药集团精方（安徽）药业股份有限公司 | Fingerprint detection method for compound Chinese medicinal decoction |
| CN114609291B (en) * | 2022-03-22 | 2023-12-26 | 国药集团精方（安徽）药业股份有限公司 | Fingerprint detection method for traditional Chinese medicine compound consistent decoction |
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