CN112557553A - Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition - Google Patents
Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition Download PDFInfo
- Publication number
- CN112557553A CN112557553A CN202011622286.9A CN202011622286A CN112557553A CN 112557553 A CN112557553 A CN 112557553A CN 202011622286 A CN202011622286 A CN 202011622286A CN 112557553 A CN112557553 A CN 112557553A
- Authority
- CN
- China
- Prior art keywords
- angelica
- fingerprint
- decoction
- decoction composition
- acetonitrile
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Landscapes
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Library & Information Science (AREA)
- Engineering & Computer Science (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a fingerprint construction method and a detection method of a Chinese angelica and six-yellow decoction composition, wherein the construction method comprises the following steps: (1) taking 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride as reference substances, and respectively preparing reference substance solutions; (2) preparing radix Angelicae sinensis, radix rehmanniae Preparata, Scutellariae radix, Coptidis rhizoma, cortex Phellodendri and radix astragali into radix Angelicae sinensis six-yellow decoction composition, and preparing the radix Angelicae sinensis six-yellow decoction composition into test solution; (3) precisely absorbing the test solution and each reference solution, injecting into a liquid chromatograph for chromatographic analysis, wherein the detection wavelength is 280 nm-320 nm, obtaining a test sample fingerprint and a reference chromatogram, and establishing a standard fingerprint of the angelica six-yellow decoction composition. The fingerprint spectrum construction method can accurately detect the standard fingerprint spectrum of the quality of the angelica sinensis Liuhuang decoction composition, and can effectively control the quality of the angelica sinensis Liuhuang decoction preparation product.
Description
Technical Field
The invention relates to the technical field of Chinese patent medicine quality detection, in particular to a fingerprint construction method and a detection method of a Chinese angelica-six-yellow decoction composition.
Background
The six-yellow decoction of angelica is from secret book of orchid Chamber of Leyuyuan codification of Lidonyuan, four gold yuan, and is known as the holy prescription for treating night sweat later, and the prescription consists of: seven medicines of Chinese angelica, radix rehmanniae, prepared rehmannia root, coptis chinensis, scutellaria baicalensis, phellodendron and astragalus membranaceus are used together, wherein the Chinese angelica, the radix rehmanniae and the prepared rehmannia root are monarch medicines, the Chinese angelica is sweet, pungent and warm in taste and has the function of nourishing blood, the radix rehmanniae is sweet, bitter and cool in taste, and the prepared rehmannia root is sweet and warm in taste and has the effect of nourishing yin; the scutellaria baicalensis is bitter and cold in taste, and can clear the upper-jiao fire, the coptis chinensis is bitter and cold in taste, and can clear the middle-jiao fire, and the phellodendron bark is bitter and cold in taste, and can purge the lower-jiao fire, so that the phellodendron bark is a ministerial drug; radix astragali is used as adjuvant drug for invigorating qi and consolidating superficial resistance. It can be used for treating night sweat due to hyperactivity of fire and yin deficiency, manifested by fever, night sweat, flushed face, dry mouth, vexation, dry lip, dry stool, yellowish or reddish urine, and red tongue with yellow fur. The party belongs to one of 100 classical famous parties in a catalog (the first batch) of ancient classical famous parties, which is prepared by the national State administration of traditional Chinese medicine and supervision and administration in 2018. The classical famous prescription is a traditional Chinese medicine compound preparation which is produced by modern industrialization based on a preparation method recorded by ancient medical records, and except for a forming process, other preparation methods are basically consistent with the records of the ancient medical records. The clinical curative effect of the classical famous prescription is exact, and the quality of the compound preparation is controlled by using a modern analysis technology, so that the stable quality of the compound preparation is ensured, and the clinical application of the classical famous prescription can be better promoted.
The traditional Chinese medicine fingerprint is an important quality analysis means for controlling traditional Chinese medicine and traditional Chinese medicine compound preparation, and is widely applied to the control of traditional Chinese medicine quality standard. At present, few researches on the fingerprint spectrum of the angelica sinensis Liuhuang decoction are carried out, and no report is made on the aspects of component analysis and quality control of the angelica sinensis Liuhuang decoction. Therefore, the method has great development value. In order to more comprehensively and effectively control the quality of the clinical medication of the angelica six-yellow decoction and ensure the safety and the curative effect of the angelica six-yellow decoction, a more advanced quality detection means needs to be adopted for the traditional Chinese medicine composition.
Disclosure of Invention
The invention aims to provide a fingerprint construction method and a detection method of a Chinese angelica six-yellow decoction composition, which can effectively control the quality of Chinese angelica six-yellow decoction preparation products.
In order to achieve the purpose, the invention adopts the following technical scheme:
a fingerprint construction method of a Chinese angelica and six-yellow decoction composition comprises the following steps:
(1) taking 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride as reference substances, and respectively preparing reference substance solutions;
(2) preparing radix Angelicae sinensis, radix rehmanniae Preparata, Scutellariae radix, Coptidis rhizoma, cortex Phellodendri and radix astragali into radix Angelicae sinensis six-yellow decoction composition, and preparing the radix Angelicae sinensis six-yellow decoction composition into test solution;
(3) precisely absorbing the test solution and each reference solution, injecting into a liquid chromatograph for chromatographic analysis, wherein the detection wavelength is 280 nm-320 nm, obtaining a test sample fingerprint and a reference chromatogram, and establishing a standard fingerprint of the angelica six-yellow decoction composition.
Further, in the step (3), the conditions for performing chromatographic analysis by the liquid chromatograph are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the number of theoretical plates is greater than or equal to 10000 calculated according to baicalin peak.
Further, the acetonitrile-0.1% phosphoric acid solution is eluted by gradient in the following order:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%;
44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
Further, the standard fingerprint contains 20 characteristic peaks which are sequentially numbered as 1-20 according to the appearance time sequence, peaks 2, 4, 5, 9, 13 and 15 in the standard fingerprint respectively correspond to retention time of chromatographic peaks of 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride, wherein the peak 13 is an S peak.
Further, the retention time of 20 characteristic peaks in the standard fingerprint spectrum is as follows from the number 1-20: 0.071, 0.159, 0.357, 0.379, 0.437, 0.482, 0.512, 0.520, 0.626, 0.824, 0.830, 0.852, 1.000, 1.035, 1.064, 1.154, 1.235, 1.278, 1.324, and 1.374.
Further, in the step (1), the 5-hydroxymethylfurfural, the chlorogenic acid, the phellodendrine hydrochloride, the ferulic acid, the baicalin and the berberine hydrochloride are respectively precisely weighed and then respectively added with 70% methanol to prepare a solution containing 10-100 μ g of solute per 1ml, which is used as a reference solution.
Further, in the step (2), 0.1g of the angelica sinensis and six-yellow soup composition is precisely weighed, 50ml of methanol with the concentration of 70% is precisely added, ultrasonic treatment is carried out, and the weight loss is compensated by the methanol with the concentration of 70%, so that the test solution is obtained.
Further, the preparation method of the angelica sinensis six-yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 4g of angelica, 4g of radix rehmanniae recen, 4g of prepared rehmannia root, 4g of scutellaria baicalensis, 4g of coptis chinensis, 4g of phellodendron amurense and 8g of astragalus membranaceus;
b. putting the weighed decoction pieces into a 2L casserole together, adding 600ml of water, soaking for 30 minutes, covering and boiling, keeping slightly boiling until the decoction is 300-400ml, and filtering with 200-mesh filter cloth while hot to obtain a filtrate;
c. concentrating the filtrate under reduced pressure, sequentially freeze drying, pulverizing and mixing to obtain the composition.
Or the preparation method of the angelica six-yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 250g of angelica, 250g of radix rehmanniae recen, 250g of prepared rehmannia root, 250g of scutellaria baicalensis, 250g of coptis chinensis, 250g of phellodendron and 500g of astragalus;
b. adding 20 times of water into the weighed decoction pieces, decocting for 60 minutes, and filtering out residues to obtain filtrate;
c. concentrating under reduced pressure to obtain fluid extract with relative density of 1.0-1.20, sequentially adding maltodextrin, drying, mixing, adding magnesium stearate, and dry-pressing to obtain the final product.
A fingerprint spectrum detection method of a Chinese angelica and six-yellow decoction composition comprises the following steps:
preparing the angelica sinensis six-yellow decoction composition to be detected into a sample solution to be detected;
injecting the sample solution to be detected into a liquid chromatograph for chromatographic analysis to generate a sample map to be detected;
the standard fingerprint spectrum in the fingerprint spectrum construction method of the angelica sinensis Liuhuang decoction composition is used as a reference spectrum, the similarity between the spectrum of the sample to be detected and the standard fingerprint spectrum is calculated, and the quality is qualified if the similarity is more than 0.9.
Further, the conditions for injecting the sample solution to be tested into the liquid chromatograph for chromatographic analysis are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the detection wavelength is 280 nm-320 nm, and the number of theoretical plates is more than or equal to 10000 calculated according to the baicalin peak;
the acetonitrile-0.1% phosphoric acid solution is eluted in a gradient manner, and the elution order is as follows:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%;
44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
The invention has the beneficial effects that:
according to the fingerprint construction method of the angelica sinensis Liuhuang decoction composition, 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride are used as reference substances, the detection wavelength of 280 nm-320 nm is used for detecting the specific components of each medicine in the composition, the sensitivity and the detection signal of each chemical component in the composition are met, and the higher level can be achieved and the detection can be carried out simultaneously.
The method uses acetonitrile-0.1% phosphoric acid solution as mobile phase to achieve good separation effect of each component in the composition, and uses 70% methanol as solvent to extract the components in the composition to generate the most chromatographic peaks.
In conclusion, the fingerprint construction method of the angelica sinensis six-yellow decoction composition can accurately detect the standard fingerprint of the quality of the angelica sinensis six-yellow decoction composition, and can effectively control the quality of the angelica sinensis six-yellow decoction preparation product.
Drawings
FIG. 1 is a 3D plot of the 200-400 nm scan spectra in the fingerprinting assay of example 3;
FIG. 2 is a chromatogram with a detection wavelength of 280nm to 320nm in example 3;
FIG. 3 is a chromatogram of each reference solution in example 3;
FIG. 4 is a chromatogram for investigation of the different mobile phase elution systems of example 4;
FIG. 5 is a chromatogram for examining different extraction solvents in example 4;
FIG. 6 is a fit graph of a chromatogram of the 10 first Angelica sinensis Liuhuang decoction composition of example 6;
FIG. 7 is the fingerprint of the Angelica sinensis Liuhuang decoction standard of example 6;
fig. 8 is a fit of chromatograms of the second angelica six-yellow decoction composition of batch 3 in example 7.
Detailed Description
The technical solution of the present invention will be further described with reference to the following embodiments.
The fingerprint construction method and the detection method of the angelica six-yellow decoction composition are used for angelica six-yellow decoction preparation products.
The angelica and six-yellow decoction is mainly used for treating yin deficiency and fire excess symptoms such as hyperthyroidism, tuberculosis, diabetes and the like in clinic. The seven medicines have the drug effects respectively, and the main pharmacological activity of the triterpenoid saponin component in the astragalus has the functions of regulating the immunity, strengthening the heart, resisting inflammation, inhibiting the synthesis of collagen and the like; iridoid glycoside in rehmannia can effectively prevent and treat diabetes and reduce blood fat, and has various effects on immune system, blood system, central nervous system, anti-aging effect and the like; when ferulic acid in radix Angelicae sinensis has anti-inflammatory effect, coptisine in rhizoma Coptidis and berberine in cortex Phellodendri belong to alkaloids, and have functions of improving glucose metabolism, lowering blood sugar and improving abnormal glucose tolerance. Baicalin, baicalein, wogonin and wogonin in Scutellariae radix have effects of resisting lipid peroxidation, inhibiting aldose reductase, and preventing and treating diabetic peripheral neuropathy and diabetic nephropathy. Therefore, based on the specific components in the angelica six-yellow decoction, the fingerprint construction method of the angelica six-yellow decoction composition adopts 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride as reference substances.
Specifically, the invention provides a fingerprint construction method of a angelica six-yellow decoction composition, which comprises the following steps:
(1) taking 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride as reference substances, and respectively preparing reference substance solutions;
(2) preparing radix Angelicae sinensis, radix rehmanniae Preparata, Scutellariae radix, Coptidis rhizoma, cortex Phellodendri and radix astragali into radix Angelicae sinensis six-yellow decoction composition, and preparing the radix Angelicae sinensis six-yellow decoction composition into test solution;
(3) precisely absorbing the test solution and each reference solution, injecting into a liquid chromatograph for chromatographic analysis, wherein the detection wavelength is 280 nm-320 nm, obtaining a test sample fingerprint and a reference chromatogram, and establishing a standard fingerprint of the angelica six-yellow decoction composition.
Because the angelica six-yellow decoction composition contains seven medicines, each medicine also contains more components and has complex chemical components. The maximum absorption wavelength of the specific components in each medicine is different, the maximum absorption wavelength of ferulic acid in angelica is 316nm, the maximum absorption wavelength of baicalin in scutellaria is 280nm, the maximum absorption wavelength of phellodendrine in phellodendron is 284nm, and the maximum absorption wavelength of berberine hydrochloride in coptis is 265 nm. The specific components of the medicines belong to alkaloids, saponins and the like, the maximum absorption wavelengths are different, and in order to meet the sensitivity and detection signals of the components and simultaneously achieve a higher level and be simultaneously measured, the detection wavelength is set to be 280-320 nm. Preferably, 290nm of detection length is selected, the separation degree of each component is good during detection, the base line is stable, and the peak shape is symmetrical.
Further, in the step (3), the conditions for performing chromatographic analysis by the liquid chromatograph are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the number of theoretical plates is greater than or equal to 10000 calculated according to baicalin peak.
The mobile phase is acetonitrile-0.1 percent phosphoric acid solution for elution, the base line of chromatographic analysis is stable, the peak shape of chromatographic peak is symmetrical, and each component has better separation effect. Specifically, acetonitrile is used as a mobile phase A, and a 0.1% phosphoric acid solution is used as a mobile phase B. Meanwhile, the column temperature is 20-30 ℃ and the flow rate is 0.2-0.3 ml/min, so that the components in the composition can be well separated. Preferably, the column temperature is 20 ℃ and the flow rate is 0.2 ml/min.
Wherein, the 0.1% phosphoric acid solution refers to a phosphoric acid aqueous solution with a volume fraction of 0.1%.
Further, the acetonitrile-0.1% phosphoric acid solution is eluted by gradient in the following order:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%;
44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
The angelica and six-yellow decoction composition contains seven medicines of angelica, radix rehmanniae recen, prepared rehmannia root, coptis root, scutellaria baicalensis, phellodendron and astragalus, and each medicine contains more components, so that the chromatographic separation difficulty is higher. Meanwhile, the invention sets more elution gradients, improves column efficiency, has certain distance between each chromatographic peak in the obtained chromatogram, can be well identified, and has proper peak shape symmetry width of the chromatographic peak.
Further, the standard fingerprint contains 20 characteristic peaks which are sequentially numbered as 1-20 according to the appearance time sequence, peaks 2, 4, 5, 9, 13 and 15 in the standard fingerprint respectively correspond to retention time of chromatographic peaks of 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride, wherein the peak 13 is an S peak. The retention time of 20 characteristic peaks in the standard fingerprint spectrum is as follows from the number 1-20: 0.071, 0.159, 0.357, 0.379, 0.437, 0.482, 0.512, 0.520, 0.626, 0.824, 0.830, 0.852, 1.000, 1.035, 1.064, 1.154, 1.235, 1.278, 1.324, and 1.374.
Specifically, the Chinese angelica and six-yellow decoction composition adopts a Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition) issued by the national pharmacopoeia committee as fingerprint similarity calculation software. And (3) introducing a similarity evaluation system based on the fingerprint chromatograms of 10 batches of test articles, selecting a chromatographic peak with stable relative retention time and good separation degree as a characteristic peak and determining the characteristic peak as a common peak, finally determining 20 characteristic peaks through comparison with a method for calculating relative retention time and relative peak area through multiple experimental studies, and automatically generating the standard fingerprint of the angelica sinensis Liuhuang decoction by using software.
Further, in the step (1), the 5-hydroxymethylfurfural, the chlorogenic acid, the phellodendrine hydrochloride, the ferulic acid, the baicalin and the berberine hydrochloride are respectively precisely weighed and then respectively added with 70% methanol to prepare a solution containing 10-100 μ g of solute per 1ml, which is used as a reference solution.
5-hydroxymethyl furfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride correspond to the specific components of the Chinese medicinal materials of the angelica six-ingredient decoction composition, and the generated chromatographic peak can be better used as a reference. Wherein, 70% methanol refers to 70% methanol aqueous solution by volume fraction. The concentration of the reference solution is limited to 10-100 μ g solute per 1ml, which is closer to the concentration of the corresponding component in the angelica six-yellow decoction composition.
Further, in the step (2), 0.1g of the angelica sinensis and six-yellow soup composition is precisely weighed, 50ml of methanol with the concentration of 70% is precisely added, ultrasonic treatment is carried out, and the weight loss is compensated by the methanol with the concentration of 70%, so that the test solution is obtained. The angelica six-yellow decoction composition is dissolved by adopting methanol with the concentration of 70 percent, the chromatographic peak of the angelica six-yellow decoction composition is the most, and the chromatographic peak is basically not lost.
Further, the preparation method of the angelica sinensis six-yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 4g of angelica, 4g of radix rehmanniae recen, 4g of prepared rehmannia root, 4g of scutellaria baicalensis, 4g of coptis chinensis, 4g of phellodendron amurense and 8g of astragalus membranaceus;
b. putting the weighed decoction pieces into a 2L casserole together, adding 600ml of water, soaking for 30 minutes, covering and boiling, keeping slightly boiling until the decoction is 300-400ml, and filtering with 200-mesh filter cloth while hot to obtain a filtrate;
c. concentrating the filtrate under reduced pressure, sequentially freeze drying, pulverizing and mixing to obtain the composition.
Or the preparation method of the angelica six-yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 250g of angelica, 250g of radix rehmanniae recen, 250g of prepared rehmannia root, 250g of scutellaria baicalensis, 250g of coptis chinensis, 250g of phellodendron and 500g of astragalus;
b. adding 20 times of water into the weighed decoction pieces, decocting for 60 minutes, and filtering out residues to obtain filtrate;
c. concentrating under reduced pressure to obtain fluid extract with relative density of 1.0-1.20, sequentially adding maltodextrin, drying, mixing, adding magnesium stearate, and dry-pressing to obtain the final product.
The preparation method of the two angelica six-yellow decoction compositions corresponds to the production process of the two angelica six-yellow decoction composition preparations. The radix Angelicae sinensis Liuhuang decoction composition obtained by the two preparation methods can be used as a test sample. It can be understood that the angelica six-yellow decoction composition obtained by other production processes can be used as a detected product.
Correspondingly, the invention provides a fingerprint spectrum detection method of the angelica six-yellow decoction composition, which comprises the following steps:
preparing the angelica sinensis six-yellow decoction composition to be detected into a sample solution to be detected;
injecting the sample solution to be detected into a liquid chromatograph for chromatographic analysis to generate a sample map to be detected;
the standard fingerprint spectrum in the fingerprint spectrum construction method of the angelica sinensis Liuhuang decoction composition is used as a reference spectrum, the similarity between the spectrum of the sample to be detected and the standard fingerprint spectrum is calculated, and the quality is qualified if the similarity is more than 0.9.
The similarity of the chromatographic fingerprint is used for judging the similarity between a sample and a standard spectrum, three common similarity calculation methods are respectively an included angle cosine method, a correlation coefficient method and an Euclidean distance method, the current similarity evaluation usually adopts a traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) provided by the State pharmacopoeia Committee to calculate the included angle cosine method, and the sample can be regarded as qualified when the general phase similarity is more than 0.9.
Further, the conditions for injecting the sample solution to be tested into the liquid chromatograph for chromatographic analysis are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the detection wavelength is 280 nm-320 nm, and the number of theoretical plates is more than or equal to 10000 calculated according to the baicalin peak;
the acetonitrile-0.1% phosphoric acid solution is eluted in a gradient manner, and the elution order is as follows:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%;
44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
The chromatographic condition can effectively separate each component in the sample solution to be detected so as to ensure accurate detection effect.
Preferably, the detection wavelength is 290 nm.
Preferably, the method for preparing the angelica six-yellow decoction composition to be detected into the sample solution to be detected comprises the following steps: precisely weighing 0.1g of the angelica and six-yolk decoction composition to be detected, precisely adding 50ml of methanol with the concentration of 70%, performing ultrasonic treatment, and complementing the weight loss by using the methanol with the concentration of 70% to obtain the sample solution to be detected.
Preferably, the preparation method of the angelica six-yellow decoction composition to be tested is the same as the preparation method of the angelica six-yellow decoction composition in the test solution. It can be understood that the angelica six-yellow decoction composition obtained by other production processes can be used as a detected product.
The invention is further illustrated by the following examples.
The reagents and apparatus involved in the following examples are as follows.
Reagent:
ferulic acid control (batch No. 110773-201614, China pharmaceutical biologicals assay research institute);
baicalin reference (batch No. 110715-201720, China institute for testing and testing biological products);
chlorogenic acid reference (batch No. 112002-201702, China institute for testing and testing biological products);
berberine hydrochloride reference (batch No. 110713-201813, China institute for testing and testing biological products of drugs);
phellodendrine hydrochloride reference (batch No. 111895-201504, China institute for testing and testing biological drugs);
5-hydroxymethylfurfural control (batch No. 111626-201610, China pharmaceutical biologicals assay institute);
the angelica six-yellow decoction composition (self-made), methanol (BCR company, USA, chromatographic purity), acetonitrile (BCR company, USA, chromatographic purity) reagent is analytically pure, and water is ultrapure water.
The instrument comprises the following steps:
waters H-class ultra high performance liquid chromatograph; waters PDA detector; an Empower workstation;
waters Acquity UPLC Cortects T3 (150X 2.1mm, 1.6 μm) chromatography column;
one-ten-thousandth analytical balance (AL104, mettler-toledo);
one part per ten million analytical balance (MS105DU, mettler-toledo corporation);
ultrasonic cleaning machines (KQ-500DE ultrasonic instruments Co., Ltd., Kunshan city);
ultra pure water systems (Merck corporation, Millipiore Synergy UV);
model TRL-05 Freeze dryer (Dalianshuangri technologies, Inc.).
EXAMPLE 1 preparation of reference solutions
Ferulic acid control solution: taking a proper amount of ferulic acid reference substance, precisely weighing, and adding 70% methanol to prepare ferulic acid solution containing 50 μ g per 1 ml.
Baicalin control solution: taking appropriate amount of baicalin control, precisely weighing, and adding 70% methanol to obtain baicalin solution containing 50 μ g of baicalin per 1 ml.
Berberine hydrochloride reference solution: taking a proper amount of berberine hydrochloride reference substance, precisely weighing, and adding 70% methanol to prepare berberine hydrochloride solution containing 50 μ g/1 ml.
Phellodendrine hydrochloride reference substance solution: taking a proper amount of phellodendrine hydrochloride reference substance, precisely weighing, and adding 70% methanol to prepare 50 μ g of phellodendrine hydrochloride solution per 1 ml.
5-hydroxymethylfurfural control solution: taking a proper amount of 5-hydroxymethylfurfural reference substance, precisely weighing, and adding 70% methanol to prepare 5-hydroxymethylfurfural solution containing 50 microgram per 1 ml.
Chlorogenic acid control solution: taking a proper amount of chlorogenic acid reference substance, precisely weighing, and adding 70% methanol to obtain chlorogenic acid solution containing 50 μ g per 1 ml.
EXAMPLE 2 preparation of test solutions
The preparation method of the angelica six-yellow decoction composition comprises two preparation methods, namely a first angelica six-yellow decoction composition and a second angelica six-yellow decoction composition are prepared by adopting the two preparation methods.
The first angelica and six-yellow decoction composition is prepared by the following steps:
a. weighing the following decoction pieces in parts by weight: 4g of angelica, 4g of radix rehmanniae recen, 4g of prepared rehmannia root, 4g of scutellaria baicalensis, 4g of coptis chinensis, 4g of phellodendron amurense and 8g of astragalus membranaceus;
b. putting the weighed decoction pieces into a 2L casserole together, adding 600ml of water, soaking for 30 minutes, covering and boiling, keeping slightly boiling until the decoction is 300-400ml, and filtering with 200-mesh filter cloth while hot to obtain a filtrate;
c. concentrating the filtrate under reduced pressure, sequentially freeze drying, pulverizing, and mixing to obtain the first radix Angelicae sinensis and six radix astragali decoction composition.
The preparation method of the second angelica and six yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 250g of angelica, 250g of radix rehmanniae recen, 250g of prepared rehmannia root, 250g of scutellaria baicalensis, 250g of coptis chinensis, 250g of phellodendron and 500g of astragalus;
b. adding 20 times of water into the weighed decoction pieces, decocting for 60 minutes, and filtering out residues to obtain filtrate;
c. concentrating under reduced pressure to obtain fluid extract with relative density of 1.0-1.20, sequentially adding maltodextrin, drying, mixing, adding magnesium stearate, and dry-pressing to obtain the second radix Angelicae sinensis and Liuhuang decoction composition.
Respectively preparing a first angelica six-yellow decoction composition and a second angelica six-yellow decoction composition into test solution, wherein the preparation method comprises the following steps:
precisely weighing 0.1g of the angelica sinensis and six-yellow soup composition, precisely adding 50ml of 70% methanol, performing ultrasonic treatment, and complementing the weight loss by 70% methanol to obtain the test solution.
Example 3 examination of chromatographic conditions of finger print
(1) Selection of detection wavelength
The sample solution prepared from the first angelicae sinensis and six-yellow decoction composition of example 2 and each reference substance solution of example 1 were precisely extracted and injected into a liquid chromatograph for chromatographic analysis. Octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile is taken as a mobile phase A, 0.1 percent phosphoric acid solution is taken as a mobile phase B, and gradient elution is carried out according to the specification in the following table; flow rate 0.2ml/ml, column temperature: at 20 ℃.
TABLE 1 fingerprint mobile phase gradient program
The full-wavelength detection is carried out on the test solution by adopting a diode array detector, the information amount of the peak at the wavelength of 290nm of the first angelica six-yellow decoction composition is larger according to the collected three-dimensional atlas (shown in figure 1) and atlases at the wavelengths of 280nm, 290nm, 300nm, 310nm and 320nm (shown in figure 2), and the comprehensive investigation and comparison show that the information amount of the peak at each wavelength is larger, so that the 290nm is the optimal detection wavelength of the fingerprint of the angelica six-yellow decoction composition.
Chromatograms (shown in fig. 3) of the reference solutions of example 1 were obtained at 290nm detection wavelength and the above chromatographic conditions, respectively, to facilitate the selection of chromatographic peak of the angelica six-yellow decoction composition.
(2) Investigation of chromatographic systems
Based on the conditions of the chromatographic analysis in the step (1), adopting 290nm detection wavelength, replacing mobile phase acetonitrile-0.1% phosphoric acid solution with acetonitrile-phosphoric acid water, methanol-phosphoric acid water, acetonitrile-formic acid water or acetonitrile-acetic acid water system, precisely sucking the test solution prepared from the first angelica six-yellow decoction composition in the example 2, injecting the test solution into a chromatograph, carrying out chromatographic analysis, and detecting fingerprint spectra as shown in a figure 4, wherein the result shows that: the first angelica six-yellow decoction composition has stable baseline in an acetonitrile-0.1% phosphoric acid water system, and the chromatographic peak type and the separation effect are obviously superior to other mobile phase systems, so that the mobile phase system is determined to be acetonitrile-0.1% phosphoric acid solution.
(3) Selection of column temperature
Based on the chromatographic analysis conditions in the step (1), the column temperature is adjusted to 30 ℃ by adopting a detection wavelength of 290nm, the sample solution prepared from the first angelica and six-yellow decoction composition in the embodiment 2 is precisely absorbed and injected into a chromatograph, and the obtained fingerprint is similar to the chromatogram obtained by the detection wavelength of 290nm in the step (1), which shows that the chromatogram with better effect can still be generated when the column temperature is 30 ℃.
EXAMPLE 4 examination of the method for preparing a test article
Based on the chromatographic analysis conditions in example 3(1), the method for preparing test solution by using different first angelica six-yellow decoction compositions with 290nm detection wavelength comprises the following steps:
taking 0.1g of the first angelica and six-yellow decoction composition, precisely weighing, respectively adding appropriate amounts of methanol, 70% methanol and water, carrying out ultrasonic treatment (power 250W and frequency 40kHz) for 30 minutes, cooling, weighing again, supplementing the lost weight with corresponding solvents, shaking up, filtering, and taking the subsequent filtrate to obtain three different test sample solutions.
Precisely sucking three different sample solutions of 1 μ l, respectively, injecting into liquid chromatograph, and measuring according to the above chromatographic conditions, the result is shown in FIG. 5. Compared with the sample prepared by 70% methanol, the sample prepared by other solvents has the defect of chromatographic peak, and the number of the extraction peak of the first angelica six-yellow decoction composition by 70% methanol is the largest, so 70% methanol is selected as the extraction solvent of the fingerprint of the angelica six-yellow decoction.
Example 5 methodological study
5.1 specificity test
Precisely sucking 1 μ l of each of the reference solution, the test solution and the 70% methanol blank solvent, injecting into a liquid chromatograph, and detecting according to the chromatographic analysis conditions in the embodiment 3(1) and the detection wavelength of 290nm, wherein the corresponding chromatographic peak appears at the corresponding position of the reference chromatogram in the result, and no corresponding peak exists on the blank sample, which indicates that the sample is negative and has no interference.
5.2 fingerprint precision test
Performing detection operation according to the chromatographic analysis conditions in the embodiment 3 and (1) and the detection wavelength of 290nm, taking S1 batches of samples, preparing a sample solution by a method for preparing the sample solution by using the first angelica sixty-yellow decoction composition in the embodiment 2, continuously injecting samples for 6 times, deriving a chromatogram, calculating the similarity by using 2012-version traditional Chinese medicine chromatographic fingerprint similarity software, and indicating that the precision of the instrument is good, wherein the result similarity is more than 0.95.
5.3 fingerprint stability test
Performing detection operation according to the conditions of chromatographic analysis in the embodiment 3 and (1) and the detection wavelength of 290nm, preparing a sample solution from S1 batches by using the method for preparing the sample solution from the first angelica sixty-yellow decoction composition in the embodiment 2, performing sample injection measurement on the prepared samples at 0 th, 4 th, 8 th, 12 th, 20 th and 24 th hours respectively, deriving a chromatogram, and calculating the similarity by using 2012-version traditional Chinese medicine chromatographic fingerprint similarity software, wherein the similarity of the results is more than 0.95, which indicates that the stability of the sample solution is good within 24 hours after the preparation is completed.
5.4 fingerprint repeatability test
Performing detection operation according to the chromatographic analysis conditions in the embodiment 3 and (1) and the detection wavelength of 290nm, taking S1 batches of samples, preparing a sample solution by a sample solution preparation method of the first angelica sixty-yellow decoction composition in the embodiment 2, feeding 6 samples respectively for determination, deriving a chromatogram, calculating similarity by using 2012-version traditional Chinese medicine chromatographic fingerprint similarity software, wherein the result similarity is more than 0.95, and the fingerprint determination method has good repeatability.
5.5 fingerprint intermediate precision test
Different experimenters perform detection operation on different dates according to the chromatographic analysis conditions in the embodiment 3 and 1 and the detection wavelength of 290nm, samples of S1 batches are taken to prepare a sample solution by a method of preparing the sample solution from the first angelica six-yellow decoction composition in the embodiment 2, 6 samples are respectively injected and measured, the similarity is calculated by deriving a chromatogram and applying 2012-version traditional Chinese medicine chromatogram fingerprint similarity software, the result similarity is all above 0.95, and the fingerprint measurement method is proved to have good intermediate precision.
The test results show that the method has the advantages of good stability, good repeatability and reproducibility, and can comprehensively reflect the integral and internal quality of the angelica six-yolk decoction composition.
Example 6 determination of common peaks and establishment of Standard fingerprints
The first angelica root-six-yolk decoction composition was prepared in 10 parts by using different batches of decoction pieces, and the sample solution was prepared by the method of preparing the sample solution from the first angelica root-six-yolk decoction composition in example 2, and the measurement was performed by using the detection wavelength of 290nm under the conditions of the chromatographic analysis in example 3 (1).
And (3) introducing the 10 batches of test solution chromatograms into 2012 versions of traditional Chinese medicine chromatogram fingerprint similarity software, determining that the common characteristic peaks are 20 by adopting a full-spectrum matching mode, taking the No. 13 baicalin chromatographic peak as an S peak, and calculating the relative retention time of the common peaks of the 10 batches of samples. The relative retention time deviation RSD of the 20 common characteristic peaks is less than 2 percent, namely:
the relative retention time RRT of the No. 1 peak is 0.071, and the RSD% is 0.01%;
the relative retention time RRT of No. 2 peak is 0.159, and RSD% is 0.02%;
the relative retention time RRT of the No. 3 peak is 0.357, and the RSD% is 0.04%;
the relative retention time RRT of the No. 4 peak is 0.379, and the RSD% is 0.05%;
the relative retention time RRT of the No. 5 peak is 0.437, and the RSD% is 0.04%;
the relative retention time RRT of the No. 6 peak is 0.482, and the RSD% is 0.03%;
the relative retention time RRT of the No. 7 peak is 0.512, and the RSD% is 0.02%;
the relative retention time RRT of the No. 8 peak is 0.520, and the RSD% is 0.02%;
the relative retention time RRT of the No. 9 peak is 0.626, and the RSD% is 0.02%;
the 10 peak relative retention time RRT is 0.824, and the RSD% is 0.04%;
relative retention time RRT of No. 11 peak is 0.830, and RSD% is 0.06%;
the relative retention time RRT of the No. 12 peak is 0.852, and the RSD% is 0.05%;
the relative retention time RRT of the No. 15 peak is 1.064, and the RSD% is 0.04%;
the relative retention time RRT of the No. 16 peak is 1.154, and the RSD% is 0.06%;
relative retention time RRT of No. 17 peak is 1.235, and RSD% is 0.05%;
relative retention time RRT of No. 18 peak is 1.278, RSD% is 0.03%;
the relative retention time RRT of the 19 th peak is 1.324, and the RSD% is 0.02%;
relative retention time RRT of No. 20 peak is 1.374, and RSD% is 0.01%;
among them, the S peak No. 13 is a chromatographic peak of the reference substance.
Fitting 10 batches of first angelica sinensis Liuhuang decoction composition samples (see figure 6) by using traditional Chinese medicine chromatography fingerprint similarity evaluation software (2012 edition), and generating an angelica sinensis Liuhuang decoction comparison fingerprint (see figure 7), namely the standard fingerprint. The sample similarity was calculated using the angle cosine method, as shown in the table below. The result shows that the similarity of 10 batches of the first angelica and six-yellow decoction composition is between 0.921 and 0.989, the similarity is higher, and the chemical components are stable.
Table 210 parts of first angelic six-yellow decoction composition similarity evaluation results
Example 7 detection of Angelica sinensis Liuhuang decoction composition by Standard fingerprint
Taking the decoction pieces in the first angelicae sinensis and rheum officinale decoction composition in example 6, 3 batches of second angelicae sinensis and rheum officinale decoction composition samples were prepared and prepared into test solution by the method in example 2. Chromatograms of 3 batches of the second angelica-six-yellow decoction composition were determined according to the conditions of the chromatography in example 3(1) and by performing the detection operation using a detection wavelength of 290 nm.
The 3 batches of chromatograms were introduced into similarity software (see fig. 8), and the standard fingerprint of example 6 was used as a satellite control, to calculate the similarity of the 3 batches of the second angelica-six-huang decoction composition, as shown in the following table. The results show that the similarity of the second angelica six-yellow decoction composition of 3 batches is more than 0.9, which shows that the quality of the three angelica six-yellow decoction preparations is stable.
TABLE 3 calculation results of the similarity of the second Dang Gui Liu Huang Tang composition
Other components and operations of a fingerprint construction method and a detection method of the angelica six-yellow decoction composition according to the embodiment of the invention are known to those skilled in the art and will not be described in detail herein.
In the description herein, references to the description of the terms "embodiment," "example," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
Claims (10)
1. A fingerprint construction method of a angelica sinensis Liuhuang decoction composition is characterized by comprising the following steps:
(1) taking 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride as reference substances, and respectively preparing reference substance solutions;
(2) preparing radix Angelicae sinensis, radix rehmanniae Preparata, Scutellariae radix, Coptidis rhizoma, cortex Phellodendri and radix astragali into radix Angelicae sinensis six-yellow decoction composition, and preparing the radix Angelicae sinensis six-yellow decoction composition into test solution;
(3) precisely absorbing the test solution and each reference solution, injecting into a liquid chromatograph for chromatographic analysis, wherein the detection wavelength is 280 nm-320 nm, obtaining a test sample fingerprint and a reference chromatogram, and establishing a standard fingerprint of the angelica six-yellow decoction composition.
2. The fingerprint construction method of the angelica sinensis six-yellow decoction composition according to claim 1, wherein in the step (3), the chromatographic analysis conditions of the liquid chromatograph are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the number of theoretical plates is greater than or equal to 10000 calculated according to baicalin peak.
3. The fingerprint construction method of the angelica six-yellow decoction composition according to claim 2, wherein the acetonitrile-0.1% phosphoric acid solution is eluted in a gradient manner, and the elution order is as follows:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%;
44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
4. The fingerprint construction method of the angelica sinensis six-ingredient decoction composition according to claim 1, wherein the standard fingerprint comprises 20 characteristic peaks, which are sequentially numbered as 1-20 according to the appearance time sequence, wherein peaks 2, 4, 5, 9, 13 and 15 in the standard fingerprint respectively correspond to retention times of chromatographic peaks of 5-hydroxymethylfurfural, chlorogenic acid, phellodendrine hydrochloride, ferulic acid, baicalin and berberine hydrochloride, and wherein peak 13 is an S peak.
5. The fingerprint construction method of the angelica six-yellow decoction composition according to claim 1, wherein the retention time of 20 characteristic peaks in the standard fingerprint is as follows from the number 1 to 20: 0.071, 0.159, 0.357, 0.379, 0.437, 0.482, 0.512, 0.520, 0.626, 0.824, 0.830, 0.852, 1.000, 1.035, 1.064, 1.154, 1.235, 1.278, 1.324, and 1.374.
6. The fingerprint construction method of the angelica sinensis six-ingredient decoction composition according to claim 1, wherein in the step (1), the 5-hydroxymethylfurfural, the chlorogenic acid, the phellodendrine hydrochloride, the ferulic acid, the baicalin and the berberine hydrochloride are precisely weighed and then respectively added with 70% methanol to prepare a solution containing 10-100 μ g of solute per 1ml, which is a reference solution.
7. The fingerprint construction method of angelica sinensis six-ingredient decoction composition according to claim 1, wherein in the step (2), 0.1g of angelica sinensis six-ingredient decoction composition is precisely weighed, 50ml of methanol with 70% concentration is precisely added, ultrasonic treatment is carried out, and the weight loss is compensated by the methanol with 70% concentration, so that the test solution is obtained.
8. The fingerprint construction method of the angelica six-yellow decoction composition according to claim 1, wherein the preparation method of the angelica six-yellow decoction composition is as follows:
a. weighing the following decoction pieces in parts by weight: 4g of angelica, 4g of radix rehmanniae recen, 4g of prepared rehmannia root, 4g of scutellaria baicalensis, 4g of coptis chinensis, 4g of phellodendron amurense and 8g of astragalus membranaceus;
b. putting the weighed decoction pieces into a 2L casserole together, adding 600ml of water, soaking for 30 minutes, covering and boiling, keeping slightly boiling until the decoction is 300-400ml, and filtering with 200-mesh filter cloth while hot to obtain a filtrate;
c. concentrating the filtrate under reduced pressure, sequentially freeze drying, pulverizing, and mixing to obtain radix Angelicae sinensis and six radix astragali decoction composition;
or the preparation method of the angelica six-yellow decoction composition comprises the following steps:
a. weighing the following decoction pieces in parts by weight: 250g of angelica, 250g of radix rehmanniae recen, 250g of prepared rehmannia root, 250g of scutellaria baicalensis, 250g of coptis chinensis, 250g of phellodendron and 500g of astragalus;
b. adding 20 times of water into the weighed decoction pieces, decocting for 60 minutes, and filtering out residues to obtain filtrate;
c. concentrating under reduced pressure to obtain fluid extract with relative density of 1.0-1.20, sequentially adding maltodextrin, drying, mixing, adding magnesium stearate, and dry-pressing to obtain the final product.
9. A fingerprint spectrum detection method of a angelica sinensis Liuhuang decoction composition is characterized by comprising the following steps:
preparing the angelica sinensis six-yellow decoction composition to be detected into a sample solution to be detected;
injecting the sample solution to be detected into a liquid chromatograph for chromatographic analysis to generate a sample map to be detected;
the angelica sinensis Liuhuang decoction composition of any one of claims 1-8, wherein the standard fingerprint in the fingerprint construction method is used as a reference, and the similarity between the sample spectrum to be tested and the standard fingerprint is calculated, and the quality is qualified if the similarity is more than 0.9.
10. The fingerprint detection method of the angelica sinensis six-yellow decoction composition according to claim 9, wherein the conditions for injecting the sample solution to be detected into the liquid chromatograph for chromatographic analysis are as follows:
octadecylsilane chemically bonded silica chromatographic column at 20-30 deg.C;
eluting with acetonitrile-0.1% phosphoric acid solution as mobile phase at flow rate of 0.2-0.3 ml/min;
the detection wavelength is 280 nm-320 nm, and the number of theoretical plates is more than or equal to 10000 calculated according to the baicalin peak;
the acetonitrile-0.1% phosphoric acid solution is eluted in a gradient manner, and the elution order is as follows:
0-18 min, acetonitrile 5% → 20%, 0.1% phosphoric acid solution 95% → 80%;
18-35 min, acetonitrile 20% → 25%, 0.1% phosphoric acid solution 80% → 75%;
35-44 min, acetonitrile 25% → 95%, 0.1% phosphoric acid solution 75% → 5%;
44-44.1 min, acetonitrile 95% → 5%, 0.1% phosphoric acid solution 5% → 95%; 44.1-50 min, 5% acetonitrile and 95% of 0.1% phosphoric acid solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011622286.9A CN112557553A (en) | 2020-12-30 | 2020-12-30 | Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011622286.9A CN112557553A (en) | 2020-12-30 | 2020-12-30 | Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112557553A true CN112557553A (en) | 2021-03-26 |
Family
ID=75034980
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011622286.9A Pending CN112557553A (en) | 2020-12-30 | 2020-12-30 | Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112557553A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113376273A (en) * | 2021-06-02 | 2021-09-10 | 国药集团精方（安徽）药业股份有限公司 | Detection method of HPLC (high performance liquid chromatography) characteristic spectrum of Qingxin lotus seed drink and application of characteristic spectrum |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109464531A (en) * | 2018-12-03 | 2019-03-15 | 湖南新汇制药股份有限公司 | A kind of preparation method of danggui liuhuang decoction |
| CN112098556A (en) * | 2020-09-21 | 2020-12-18 | 山东宏济堂制药集团股份有限公司 | Detection method of angelica sinensis Liuhuang decoction |
-
2020
- 2020-12-30 CN CN202011622286.9A patent/CN112557553A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109464531A (en) * | 2018-12-03 | 2019-03-15 | 湖南新汇制药股份有限公司 | A kind of preparation method of danggui liuhuang decoction |
| CN112098556A (en) * | 2020-09-21 | 2020-12-18 | 山东宏济堂制药集团股份有限公司 | Detection method of angelica sinensis Liuhuang decoction |
Non-Patent Citations (2)
| Title |
|---|
| 周德生 等: "《中医膏方临床应用指南》", 31 October 2015, 山西科学技术出版社 * |
| 宿莹: ""当归六黄汤物质基准的研究"", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113376273A (en) * | 2021-06-02 | 2021-09-10 | 国药集团精方（安徽）药业股份有限公司 | Detection method of HPLC (high performance liquid chromatography) characteristic spectrum of Qingxin lotus seed drink and application of characteristic spectrum |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113049724B (en) | Fingerprint construction method and detection method of cassia twig, chinese herbaceous peony and rhizoma anemarrhenae decoction composition | |
| CN103330758A (en) | Peony and liquorice soup formula granule, preparation method and detection method of peony and liquorice soup formula granule | |
| CN115718145A (en) | Method for measuring fingerprint spectrum of traditional Chinese medicine composition | |
| CN106290599B (en) | Content determination method of traditional Chinese medicine composition | |
| CN114200046A (en) | Construction method and detection method of Juanbi decoction composition fingerprint spectrum and application | |
| CN113063885B (en) | Composition for preparing Baoyuan decoction, baoyuan decoction product and fingerprint spectrum measuring and quality detecting method thereof | |
| CN113156019B (en) | Method for detecting multi-component content of liquorice heart-fire-purging decoction | |
| CN112557553A (en) | Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition | |
| CN110568108B (en) | Multi-component content determination method of Ganfule preparation | |
| CN108226325A (en) | Roripa montana gives birth to the method for building up of arteries and veins oral liquid composition finger-print | |
| CN115267008B (en) | Construction method of characteristic spectrum and comparison spectrum of bamboo juice pinellia ternate and distinguishing method of different processed products of pinellia ternate | |
| CN114910576B (en) | Method for detecting aconite monoester type alkaloid component in cassia twig, chinese herbaceous peony and rhizoma anemarrhenae soup | |
| CN113759056B (en) | Characteristic spectrum of Chinese lobelia and preparation thereof and construction method thereof | |
| CN104458954B (en) | A kind of dodder formulation granule finger printing and method for building up thereof | |
| CN109521122B (en) | Preparation method of fingerprint of traditional Chinese medicine preparation for treating functional dyspepsia | |
| CN114660218A (en) | Medicine composition containing 'Qingshanjuantong decoction' and detection method | |
| CN113341007A (en) | Method for measuring contents of multiple components in whole Chinese date seed nerve-soothing capsule based on HPLC (high performance liquid chromatography) characteristic spectrum | |
| CN111562324A (en) | Method for detecting content of multiple index components of medicinal preparation | |
| CN114689708B (en) | Long Guican Qijiu fingerprint detection method | |
| CN117571899B (en) | Quality control method for Chinese medicinal composition containing radix Angelicae sinensis and its application | |
| CN113984915B (en) | Method for detecting characteristic spectrum of four seasons Sanhuang pills | |
| CN115372517B (en) | Construction and application of UPLC characteristic spectrum of immature bitter orange, longstamen onion bulb and cassia twig decoction | |
| CN113759055B (en) | Method for establishing characteristic spectrum of rhizoma dioscoreae septemlobae test sample | |
| CN111474276B (en) | Quality control method of yang invigorating tablet preparation | |
| CN116223656B (en) | Detection method of dampness resolving and toxin resolving composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210326 |
|
| RJ01 | Rejection of invention patent application after publication |