CN111024877A - Method for detecting traditional Chinese medicine components in kidney-tonifying and bone-strengthening pill - Google Patents
Method for detecting traditional Chinese medicine components in kidney-tonifying and bone-strengthening pill Download PDFInfo
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Abstract
The invention discloses a method for detecting traditional Chinese medicine components in a kidney-tonifying and bone-strengthening pill, which belongs to the technical field of traditional Chinese medicine detection, wherein the method comprises the steps of observing under a microscope, primarily judging main medicine taste, grouping, carrying out thin-layer identification for qualitative judgment, then carrying out quantitative analysis on morroniside and loganin, and taking the total content of morroniside and loganin as a judgment basis; it is used as the basis for judging the qualification of the kidney-tonifying and bone-strengthening pill according to the qualitative and quantitative conformity of the traditional Chinese medicine components. The beneficial effects are that: the invention can carry out comprehensive judgment by a plurality of detection methods, can quickly judge the accuracy of the medicine components in a one-plate multi-detection mode, and simultaneously carries out quantitative analysis on specific indexes in the medicine to judge whether the specific indexes meet the set indexes. The method is specially used for detecting the kidney-tonifying and bone-strengthening pills, and the idea of one plate for multiple detections disclosed in the patent also provides a corresponding technical idea for detecting other traditional Chinese medicine components.
Description
Technical Field
The invention relates to a detection method of traditional Chinese medicine components, in particular to a detection method of traditional Chinese medicine components in a kidney-tonifying and bone-strengthening pill, and belongs to the technical field of traditional Chinese medicine detection.
Background
In the prior art, a traditional Chinese medicine pill for tonifying the kidney and strengthening bones is prepared from the following components in parts by weight: 100 parts of dogwood; 100 parts of eucommia ulmoides; 200 parts of wolfberry fruit; 100 parts of fructus psoraleae; 100 parts of red peony root; 100 parts of radix rehmanniae; 100 parts of prepared rhizome of rehmannia; 200 parts of teasel roots; 100 parts of clematis root; 100 parts of angelica; 300 parts of astragalus; 100 parts of white peony root; 100 parts of earthworm; 40 parts of fructus amomi.
The fourteen traditional Chinese medicine components are prepared by mixing and decocting five Chinese medicines of radix rehmanniae, radix rehmanniae preparata, teasel root, red paeony root and clematis root twice, adding 8 times of weight of water for 2 hours for the first time, adding 5 times of weight of water for 1 hour for the second time, mixing decoctions, filtering, concentrating the filtrate to thick paste with the relative density of 1.16-1.18 (70-80 ℃), crushing the rest nine Chinese medicines into fine powder, uniformly mixing the fine powder with the thick paste, drying for 5 hours at 100 ℃, crushing, sieving by a sieve of 80-100 meshes, and pelleting with water.
The product is a concentrated pill from yellow brown to brown; fragrant and bitter.
The functions and indications are as follows: tonify kidney, strengthen bone and strengthen body. Can be used for treating lumbago, skelalgia, arthralgia, soreness of waist and knees, and myasthenia of limbs due to liver and kidney deficiency and weak tendons and bones fistula.
The application and dosage are as follows: it is administered orally with warm water 5g each time and 3 times a day.
Due to the limitation of factors such as the growth environment, the year, the variety and the like of the traditional Chinese medicinal materials, the change of effective components in the traditional Chinese medicinal materials is large, so that the qualitative and quantitative analysis of main components in the traditional Chinese medicinal materials is particularly important. In the prior art, a great deal of records are recorded for researching, purifying and quantitatively analyzing the category of the active ingredients in a specific traditional Chinese medicinal material, but the technical scheme for researching the ingredients of the patent medicines is relatively less, and particularly, the research on the ingredients of the patent medicines after multi-ingredient mixing is less, because when a plurality of ingredients are mixed together, the ingredients and the active ingredients of the patent medicines can mutually influence each other, so that the detection results can mutually interfere, and the final index judgment is influenced; if each active ingredient is used as a single index for detection, the detection is tedious and the workload is large, so how to classify a plurality of different drugs, determine the types of the drugs by fewer detection steps, and even determine whether the content of the active ingredients meets the standard, is a technical problem to be solved in the field.
Disclosure of Invention
The invention aims to provide a method for detecting traditional Chinese medicine components in a kidney-tonifying and bone-strengthening pill, which comprehensively judges the accuracy of medicine components by a plurality of detection methods, and simultaneously carries out quantitative analysis on specific indexes in the medicine to judge whether the specific indexes meet the set indexes.
The invention aims to realize the detection method of the traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill, which comprises the following steps:
(1) grinding the tested patent medicine into powder, observing under a microscope, and primarily judging whether the tested patent medicine contains dogwood, eucommia bark, medlar, malaytea scurfpea fruit, angelica, astragalus, white paeony root, earthworm and amomum fruit;
(2) identifying a thin layer;
(2.1) taking the powder of the tested finished medicine, adding methanol, heating and refluxing, concentrating, then adding the concentrated solution on a neutral alumina column, eluting with methanol, removing the solution and evaporating to dryness, dissolving the residue in water, extracting with water-saturated n-butyl alcohol, evaporating to dryness, adding methanol for dissolution, and preparing a test solution; preparing control medicinal materials of dogwood, astragalus, white paeony root and red paeony root into a control medicinal material solution by the same method in the section; adding methanol into ursolic acid, astragaloside IV and penoniflorin as control solutions; sucking the test solution, the reference solution and the reference solution, spotting on the same silica gel G thin layer plate, performing chromatographic analysis, and observing the consistency of the characteristic spots of the chromatogram of the test solution and the chromatograms of the reference solution and the reference solution;
(2.2) taking the powder of the to-be-tested patent medicine, adding methanol, carrying out ultrasonic treatment and filtering, and concentrating the filtrate to be used as a test solution; preparing radix Angelicae sinensis and fructus Psoraleae in the same way to obtain control medicinal solution; taking ligustilide, psoralen and isopsoralen, and adding methanol respectively to obtain reference solutions; sucking the test solution, the reference medicinal material solution and the reference solution, spotting on the same silica gel G thin-layer plate, performing fluorescence chromatography, and observing the consistency of the characteristic spots of the chromatogram of the test solution, the chromatogram of the reference medicinal material solution and the chromatogram of the reference solution;
(2.3) taking the powder of the tested patent medicine, adding water, heating and refluxing, filtering and concentrating, shaking and extracting by using ethyl acetate, and concentrating again to obtain a test solution I; taking the powder of the tested patent medicine, adding ethyl acetate, filtering after ultrasonic treatment, and concentrating the filtrate to be used as a test solution II; preparing a fructus lycii reference medicinal material into a reference medicinal material solution according to the preparation method of the test solution I; preparing a reference medicinal material solution from eucommia ulmoides according to the preparation method of the test solution II; sucking a test solution I, a test solution II and a reference medicinal material solution, respectively dropping the test solution I, the test solution II and the reference medicinal material solution on the same silica gel G thin-layer plate, performing fluorescence chromatographic analysis by taking toluene-ethyl acetate-formic acid as a developing agent, and observing the consistency of characteristic spots of the chromatogram of the test solution and the chromatogram of the reference medicinal material solution;
(3) quantitative analysis of morroniside and loganin: performing HPLC analysis at 240nm by gradient elution with C18 chromatographic column and acetonitrile as mobile phase A and phosphoric acid solution with mass content of 0.3% as mobile phase B; acquiring peak areas of morroniside and loganin, and calculating the contents of morroniside and loganin according to the following regression equation:
morroniside Y-16194X +4280 regression coefficient 0.9997
Loganin Y-16186X +9019 regression coefficient 1
Wherein, Y is peak area, X is content, unit mug/mL;
calculating the sum of the contents of morroniside and loganin in the Chinese medicinal composition according to the value of X, and determining that the total amount of morroniside and loganin in the Chinese medicinal composition is not less than 0.56 mg/g.
(4) According to the qualitative and quantitative conformity of the traditional Chinese medicine components, the method is used as the basis for judging the qualification of the kidney-tonifying and bone-strengthening pill.
The invention carries out preliminary judgment by adding the medicine characteristics of the powder to obtain a preliminary judgment result, and then analyzes the correspondence of each substance according to the chromatographic contrast, thereby further accurately judging whether the kidney-tonifying and bone-strengthening pill contains the medicine flavor and the main chemical components thereof specified by the formula; meanwhile, whether the medicine meets the standard or not is determined by measuring the total content of the morroniside and the loganin. The accuracy of the medicine components can be rapidly judged by a mode of one plate and multiple tests, and meanwhile, the specific indexes in the medicine are quantitatively analyzed to judge whether the specific indexes meet the set indexes. In view of the complexity of the components of the traditional Chinese medicine, the invention determines that the effective components and the content of the traditional Chinese medicine cannot be completely judged, the invention can judge the main monarch drug and ministerial drug components in the kidney-tonifying and bone-strengthening pill, can determine the judgment standard of the kidney-tonifying and bone-strengthening pill by the index, can evaluate the kidney-tonifying and bone-strengthening pills produced by different medicine manufacturers by the standard, is specially used for detecting the kidney-tonifying and bone-strengthening pills, and provides corresponding technical ideas for testing other traditional Chinese medicine components.
Specifically, in step (1), when observed under a microscope:
the surface of the epidermal cell of the pericarp is polygonal or rectangular, the diameter is 16-30 mu m, the vertical peripheral wall is thickened in a bead shape, the outer flat peripheral wall is thickened in a granular cutin shape, the cell cavity contains light orange yellow substances, the mesocarp cell is orange brown and is more shrunken, and the pericarp cell is primarily judged to contain the dogwood; the rubber threads are stripped or twisted into balls, the surface of the rubber threads is granular, and the rubber threads are primarily judged to contain eucommia; the surface of the pericarp stone cell is irregular and polygonal, the wall thickness is thick, the wavelike bending is realized, the striations are clear, and the pericarp stone cell is primarily judged to contain the medlar; the fibers are bundled or scattered, the diameter is 8-30 mu m, the wall thickness is thick, longitudinal cracks are formed on the surface, the primary wall and the secondary wall are separated, the two ends of the primary wall and the secondary wall are always broken into whisker shapes or flat sections, and the astragalus membranaceus is preliminarily judged to be contained; the phloem parenchyma cells are spindle-shaped, the wall is slightly thick, the surface has extremely fine obliquely staggered textures, the transverse septa of the phloem parenchyma cells can be seen sometimes, and the Chinese angelica is preliminarily judged; calcium oxalate clusters with the diameter of 11-35 mu m exist in parenchymal cells and are arranged in rows, or one cell contains a plurality of clusters, and the cells are preliminarily judged to contain white paeony roots; longitudinal furrows are observed on the side surface of the gridlike cells, 1 bright band is positioned at the edge close to the upper side, the top surface is polygonal, the cell cavity is extremely small, the pore furrows are fine, and the gridlike cells are preliminarily judged to contain the fructus psoraleae; the endothelial sclereid cells are reddish brown or yellowish brown, the surfaces of the endothelial sclereid cells are polygonal, the wall thicknesses of the endothelial sclereid cells are not lignified, the intracellular cavities contain siliceous blocks, the sections of the endothelial sclereid cells are 1 row of grid cells, the inner walls and the side walls of the endothelial sclereid cells are extremely thick, the intracellular cavities are arranged on the outer sides, and the endothelial sclereid blocks are preliminarily judged to contain fructus amomi; the epidermal cells are brownish yellow, the cell boundary is not obvious, dark brown pigment particles are distributed, and the earthworm is initially judged to be contained.
The specific method of the step (2.1) is as follows:
taking 5g of test sample powder, adding 50mL of methanol, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 15mL, adding the filtrate onto a 5g neutral alumina column with a 100-120 mesh sieve, eluting with 100mL of 40% methanol aqueous solution, collecting the eluent, evaporating to dryness, dissolving the residue in 30mL of water, extracting for 2 times with water-saturated n-butanol by shaking for 20mL each time, combining the n-butanol solution, evaporating to dryness, and dissolving the residue in 1mL of methanol to obtain a test sample solution; preparing control medicinal materials of Corni fructus, radix astragali, radix Paeoniae alba, and radix Paeoniae Rubra each 1g, and making into control medicinal solution respectively by the same method; adding methanol into ursolic acid, astragaloside IV and penoniflorin as reference substances to obtain 1mg solution per 1mL as reference substance solution; in the thin layer chromatography test, sucking 5 μ L of the sample solution, 2 μ L of the reference solution and 2 μ L of the reference solution, respectively dropping on the same silica gel G thin layer plate, developing with the lower layer solution of dichloromethane-methanol-water solution at volume ratio of 13: 6: 2 as developing agent, taking out, air drying, spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and observing the consistency of the characteristic spots of the chromatogram of the sample solution and the chromatogram of the reference solution and the reference solution.
The specific method of the step (2.2) is as follows:
taking 2g of test sample powder, adding 30mL of methanol, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to be used as a test sample solution; preparing 1g of radix Angelicae sinensis and fructus Psoraleae control medicinal materials respectively, and making into control medicinal solution respectively; then taking ligustilide, psoralen and isopsoralen as reference substances, and adding methanol to obtain 1mg solution per 1mL as reference substance solution; when performing thin layer chromatography, sucking 2 μ L of each of the six solutions, respectively dropping on the same silica gel G thin layer plate, developing with n-hexane-ethyl acetate solution at volume ratio of 7: 2 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color.
The specific method of the step (2.3) is as follows:
taking 5g of test sample powder, adding 50mL of water, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 20mL, extracting with ethyl acetate by shaking for 2 times, 15mL each time, combining ethyl acetate solutions, and concentrating to 1mL to obtain a test sample solution I; taking 2g of test sample powder, adding 30mL of ethyl acetate, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to obtain a test sample solution II; taking 1g of fructus Lycii reference material, and making into reference material solution according to the preparation method of the test solution I; taking 1g of eucommia ulmoides as a reference medicinal material, and preparing a reference medicinal material solution according to a preparation method of a test solution II; when performing thin layer chromatography, sucking 5 μ L of sample solution I, 5 μ L of sample solution II and 2 μ L of control solution, respectively dropping on the same silica gel G thin layer plate, developing with toluene-ethyl acetate-formic acid solution at volume ratio of 15: 8: 1.5 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color.
The specific method for carrying out the quantitative analysis of the morroniside and the loganin in the step (3) is as follows:
taking 5g of test sample powder, precisely weighing, placing in a conical flask with a plug, precisely adding 50mL of methanol, and weighing; ultrasonic processing with power of 300W and frequency of 50kHz for 30 minutes, cooling, weighing, supplementing the weight loss with methanol, shaking up, and filtering; respectively and precisely sucking 10 mu L of test solution, injecting into a liquid chromatograph, and carrying out chromatography with Agilent Zorbax SB-C18 chromatographic column; taking acetonitrile as a mobile phase A, taking a phosphoric acid solution with the weight content of 0.3% as a mobile phase B, and carrying out gradient elution; the flow rate was 1.0mL per minute; under the column temperature of 30 ℃, the detection wavelength is 240nm, the peak areas of the morroniside and the loganin are obtained, and then calculation is carried out.
Drawings
Fig. 1 is a microscopic characteristic diagram of various medicinal materials, wherein 1 a: cornus officinalis pericarp epidermal cell surface (cornus officinalis); 1 b: cornus officinalis pericarp epidermic cell lateral view (cornus officinalis); 2: dogwood mesocarp cells (dogwood fruit); 3: rubber silk (eucommia); 4: fiber bundle (astragalus); 5: pericarp stone cells (eucommia ulmoides); 6: phloem parenchyma cells (angelica); 7: calcium oxalate clusters (white peony); 8 a: pericarp grid cell lateral view (psoralea corylifolia); 8 b: pericarp grid cell apical view (psoralea corylifolia); 9 a: endothelial pericarp pachytene surface appearance (amomum villosum); 9 b: the section of the endothelial pericarp with thick wall cells (fructus Amomi); 10: epidermal cells (earthworms).
FIG. 2 is a thin layer chromatogram corresponding to step (2.1); in the figures, 1, 12, 13: test articles (lot numbers: 180821, 181203, 190425); 2: a dogwood negative sample; 3: dogwood reference medicinal material; 4: an ursolic acid control; 5: negative control of radix astragali; 6: radix astragali as reference material; 7: astragaloside IV reference substance; 8: paeoniflorin negative control; 9: radix Paeoniae alba as reference material; 10: radix Paeoniae Rubra reference material; 11: paeoniflorin control.
FIG. 3 is a thin layer chromatogram corresponding to step (2.2); in the figures, 1-3: test articles (lot numbers: 180821, 181203, 190425); 4: negative control of fructus Psoraleae; 5: fructus Psoraleae as reference material; 6: psoralen reference; 7: isopsoralen control; 8: angelica negative control; 9: radix Angelicae sinensis as reference material; 10: ligustilide reference substance.
FIG. 4 is a thin layer chromatogram corresponding to step (2.3); in the figures, 1-3: test article I (lot numbers 180821, 181203, 190425; corresponding to test article solution I); 4: wolfberry fruit negative control; 5: fructus Lycii as reference material; 6-8: sample II (lot numbers: 180821, 181203, 190425; corresponding to sample solution II); 9: negative control of eucommia ulmoides; 10: eucommia ulmoides as reference medicinal material.
FIG. 5 is a morroniside and loganin specificity chromatogram.
Detailed Description
The detection method of the traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill is carried out according to the following method, and the steps involved in the method can be interchanged according to the actual situation.
(1) Grinding the tested patent medicine into powder, observing under a microscope, and primarily judging whether the tested patent medicine contains dogwood, eucommia bark, medlar, malaytea scurfpea fruit, angelica, astragalus, white paeony root, earthworm and amomum fruit; when observed under a microscope, as shown in fig. 1:
the surface of the epidermal cell of the pericarp is polygonal or rectangular, the diameter is 16-30 mu m, the vertical peripheral wall is thickened in a bead shape, the outer flat peripheral wall is thickened in a granular cutin shape, the cell cavity contains light orange yellow substances, the mesocarp cell is orange brown and is more shrunken, and the pericarp cell is primarily judged to contain the dogwood; the rubber threads are stripped or twisted into balls, the surface of the rubber threads is granular, and the rubber threads are primarily judged to contain eucommia; the surface of the pericarp stone cell is irregular and polygonal, the wall thickness is thick, the wavelike bending is realized, the striations are clear, and the pericarp stone cell is primarily judged to contain the medlar; the fibers are bundled or scattered, the diameter is 8-30 mu m, the wall thickness is thick, longitudinal cracks are formed on the surface, the primary wall and the secondary wall are separated, the two ends of the primary wall and the secondary wall are always broken into whisker shapes or flat sections, and the astragalus membranaceus is preliminarily judged to be contained; the phloem parenchyma cells are spindle-shaped, the wall is slightly thick, the surface has extremely fine obliquely staggered textures, the transverse septa of the phloem parenchyma cells can be seen sometimes, and the Chinese angelica is preliminarily judged; calcium oxalate clusters with the diameter of 11-35 mu m exist in parenchymal cells and are arranged in rows, or one cell contains a plurality of clusters, and the cells are preliminarily judged to contain white paeony roots; longitudinal furrows are observed on the side surface of the gridlike cells, 1 bright band is positioned at the edge close to the upper side, the top surface is polygonal, the cell cavity is extremely small, the pore furrows are fine, and the gridlike cells are preliminarily judged to contain the fructus psoraleae; the endothelial sclereid cells are reddish brown or yellowish brown, the surfaces of the endothelial sclereid cells are polygonal, the wall thicknesses of the endothelial sclereid cells are not lignified, the intracellular cavities contain siliceous blocks, the sections of the endothelial sclereid cells are 1 row of grid cells, the inner walls and the side walls of the endothelial sclereid cells are extremely thick, the intracellular cavities are arranged on the outer sides, and the endothelial sclereid blocks are preliminarily judged to contain fructus amomi; the epidermal cells are brownish yellow, the cell boundary is not obvious, dark brown pigment particles are distributed, and the earthworm is initially judged to be contained.
(2) Identifying a thin layer;
step (2.1), the specific method is as follows: taking 5g of test sample powder, adding 50mL of methanol, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 15mL, adding the filtrate onto a 5g neutral alumina column with a 100-120 mesh sieve, eluting with 100mL of 40% methanol aqueous solution, collecting the eluent, evaporating to dryness, dissolving the residue in 30mL of water, extracting for 2 times with water-saturated n-butanol by shaking for 20mL each time, combining the n-butanol solution, evaporating to dryness, and dissolving the residue in 1mL of methanol to obtain a test sample solution; preparing control medicinal materials of Corni fructus, radix astragali, radix Paeoniae alba, and radix Paeoniae Rubra each 1g, and making into control medicinal solution respectively by the same method; adding methanol into ursolic acid, astragaloside IV and penoniflorin as reference substances to obtain 1mg solution per 1mL as reference substance solution; in the thin layer chromatography test, sucking 5 μ L of the sample solution, 2 μ L of the reference solution and 2 μ L of the reference solution, respectively dropping on the same silica gel G thin layer plate, developing with the lower layer solution of dichloromethane-methanol-water solution at volume ratio of 13: 6: 2 as developing agent, taking out, air drying, spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and observing the consistency of the characteristic spots of the chromatogram of the sample solution and the chromatogram of the reference solution and the reference solution. As shown in fig. 2.
And (2.2) specifically comprising the following steps:
taking 2g of test sample powder, adding 30mL of methanol, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to be used as a test sample solution; preparing 1g of radix Angelicae sinensis and fructus Psoraleae control medicinal materials respectively, and making into control medicinal solution respectively; then taking ligustilide, psoralen and isopsoralen as reference substances, and adding methanol to obtain 1mg solution per 1mL as reference substance solution; when performing thin layer chromatography, sucking 2 μ L of each of the six solutions, respectively dropping on the same silica gel G thin layer plate, developing with n-hexane-ethyl acetate solution at volume ratio of 7: 2 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color. As shown in fig. 3.
And (2.3), specifically comprising the following steps:
taking 5g of test sample powder, adding 50mL of water, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 20mL, extracting with ethyl acetate by shaking for 2 times, 15mL each time, combining ethyl acetate solutions, and concentrating to 1mL to obtain a test sample solution I; taking 2g of test sample powder, adding 30mL of ethyl acetate, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to obtain a test sample solution II; taking 1g of fructus Lycii reference material, and making into reference material solution according to the preparation method of the test solution I; taking 1g of eucommia ulmoides as a reference medicinal material, and preparing a reference medicinal material solution according to a preparation method of a test solution II; when performing thin layer chromatography, sucking 5 μ L of sample solution I, 5 μ L of sample solution II and 2 μ L of control solution, respectively dropping on the same silica gel G thin layer plate, developing with toluene-ethyl acetate-formic acid solution at volume ratio of 15: 8: 1.5 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color. As shown in fig. 4.
When thin layer identification is performed in the steps (2.1), (2.2) and (2.3), the batch number of the sample: 180821, 181203, 190425.
In the step (2.1), dogwood, astragalus, white paeony root and red paeony root are taken as reference medicinal materials, ursolic acid, astragaloside and paeoniflorin are taken as reference substances, one-plate multi-test identification is carried out, spots with the same color are displayed on positions corresponding to the chromatograms of the reference medicinal materials and the reference substances in the chromatogram of the test solution, the negative control is not interfered, and the chromatogram is shown in figure 2.
In the step (2.2), angelica and psoralea are used as reference medicinal materials, ligustilide, psoralen and isopsoralen are used as reference substances, one-plate multi-test identification is carried out, fluorescent spots with the same color are shown in the chromatogram of the test sample at the positions corresponding to the chromatograms of the reference medicinal materials and the reference substances, the negative control is free of interference, and the detailed picture is shown in figure 3.
In the step (2.3), wolfberry fruit and eucommia bark are used as reference medicinal materials to perform one-plate multi-test identification, fluorescent spots with the same color are displayed on the positions, corresponding to the chromatogram of the reference medicinal material, in the chromatogram of the test sample, the negative control is free of interference, and detailed picture 4 is shown in detail.
(3) Quantitative analysis of morroniside and loganin: taking 5g of test sample powder, precisely weighing, placing in a conical flask with a plug, precisely adding 50mL of methanol, and weighing; ultrasonic processing with power of 300W and frequency of 50kHz for 30 minutes, cooling, weighing, supplementing the weight loss with methanol, shaking up, and filtering; respectively and precisely sucking 10 mu L of test solution, injecting into a liquid chromatograph, and carrying out chromatography with Agilent Zorbax SB-C18 chromatographic column; taking acetonitrile as a mobile phase A, taking a phosphoric acid solution with the weight content of 0.3% as a mobile phase B, and carrying out gradient elution; the flow rate was 1.0mL per minute; under the column temperature of 30 ℃, the detection wavelength is 240nm, the peak areas of the morroniside and loganin are obtained, and the contents of the morroniside and loganin are calculated according to the following regression equation:
morroniside Y-16194X +4280 regression coefficient 0.9997
Loganin Y-16186X +9019 regression coefficient 1
Wherein, Y is peak area, X is content, unit mug/mL;
and calculating the content sum of the morroniside and loganin in the finished medicine according to the value of X, wherein the judgment basis is that the total amount of the morroniside and loganin in the finished medicine is not less than 0.56 mg/g.
When gradient elution was performed, it was performed as in table 1:
TABLE 1 gradient elution Table
| Time (minutes) | A(％) | B(％) |
| 0～20 | 7 | 93 |
| 20～60 | 7→20 | 93→80 |
| 60～70 | 20 | 80 |
| 70～75 | 20→7 | 80→93 |
| 75～80 | 7 | 93 |
Taking morroniside and loganin as reference substances, precisely weighing, and adding methanol to obtain a mixed solution containing morroniside 20 μ g and loganin 50 μ g per 1 mL.
More specifically, the content of morroniside and loganin in the monarch drug cornus is determined by high performance liquid chromatography.
The instrument comprises the following steps: agilent1260 high performance liquid chromatograph, DAD detector; mettler Toledo XS205 DU analytical balance, Mettler Toledo corporation, Mettler Toledo; KH-400KDE model high power digital control ultrasonic cleaner of Kunshan Seaman ultrasonic instrument Limited; an OGH60 electric heating constant temperature drying oven of Saimei Fei company; Milli-Q ultrapure water instrument, Millipop Milli; byjek BT-150 multifunctional crusher (550W, 50-60 Hz).
Sample and reagent: kidney-tonifying and bone-strengthening pill samples (specification: 45 g/box, Baohui county traditional Chinese medicine hospital, batch numbers: 181119, 181203 and 181218); kidney-tonifying and bone-strengthening pill negative sample (lack of pulp of dogwood fruit, baying ying county traditional Chinese medical hospital, batch number: 181203); the morroniside reference substance (batch No. 111998-201602, content: 96.3%) and the loganin reference substance (batch No. 111640-201808, content: 99%) were purchased from China food and drug testing research institute; acetonitrile is chromatographically pure (Beijing Dima technical Co., Ltd.); methanol, ethanol and phosphoric acid are all analytically pure (chemical reagents of national drug group, Ltd.); the water is purified water.
Accuracy, precision, specificity, linear range, detection limit, quantification limit, etc. are examined.
First, accuracy
2.5g of sample (batch No. 181203) powder (sieved by a No. four sieve) is precisely weighed, 25mL of mixed reference substance solution containing 17.64 mu g of morroniside and 45.84 mu g of loganin is precisely added, 6 parts of test solution are prepared, the content of each component to be tested is measured, and the sample adding recovery rate is calculated, and the result is shown in Table 2.
TABLE 2 sample recovery test results
As can be seen from the above table, the method is accurate.
Second, precision
A sample (lot No. 181203) was taken as a powder, and 6 test solutions were prepared in parallel according to a standard method and the contents were calculated, whereby the contents of morroniside and loganin were 0.9% and 0.7%, respectively, in terms of RSD. In addition, the relative deviation of results of different inspectors, different brands of instruments and different dates is considered and is within 2.0%. As a result, the method is excellent in precision.
Third, specialization
Samples were taken, respectively, as powder (lot # 181203) and as powder (lot # 181203), prepared and injected according to standard methods, and chromatograms were recorded, as shown in FIG. 5. As a result, the chromatographic peaks of morroniside and loganin are detected in the sample, the separation degree of the chromatographic peaks from adjacent peaks is more than 1.5, and corresponding chromatographic peaks are not detected in a negative sample, which indicates that the negative sample is not interfered, so that the method has good specificity.
Four, linear range
Taking appropriate amount of morroniside and loganin reference substances, diluting with methanol to obtain 6 mixed reference substance solutions with different concentrations, wherein the morroniside concentrations are 2.94 μ g/mL, 5.87 μ g/mL, 11.75 μ g/mL, 29.37 μ g/mL, 46.99 μ g/mL, and 58.74 μ g/mL respectively; the loganin concentrations were 15.29. mu.g/mL, 30.57. mu.g/mL, 61.14. mu.g/mL, 152.85. mu.g/mL, 244.56. mu.g/mL, and 305.70. mu.g/mL, respectively. 10. mu.L of each of the above solutions was taken, injected into a liquid chromatograph, and a chromatogram was recorded. A standard curve was drawn with the concentration (. mu.g/mL) of each control as the abscissa X and the peak area as the ordinate Y. The linear equation, linear range and correlation coefficient are shown in table 3. The experimental result shows that the concentrations of the morroniside and the loganin have good linear relation with the peak area. Through the linear relation, the contents of the morroniside and the loganin and the sum of the contents can be calculated.
TABLE 3 results of linear relationship examination
| Component to be measured | Regression equation | r | Linear Range (μ g/mL) |
| Morroniside | Y＝16194X+4280 | 0.9997 | 2.94～58.74 |
| Loganin A. Loganin A | Y＝16186X+9019 | 1 | 15.29～305.70 |
Fifthly, detection limit and quantification limit
Taking appropriate amount of morroniside and loganin reference substances, diluting with methanol to obtain solution with appropriate concentration, analyzing 10 μ L sample injection with S/N of 3 as index to obtain morroniside sample injection concentration of 0.29 μ g/mL, with detection limit of 2.9 × 10-3Mu g; the loganin injection concentration is 0.31 mug/mL, namely the detection limit is 3.1 multiplied by 10-3Mu g; the S/N ratio is 10, the concentration of the morroniside sample is 0.98 mug/mL, namely the limit of quantitation is 9.8 multiplied by 10-3Mu g; the concentration of the loganin sample is 1.02 mu g/mL, namely the limit of quantitation is 10.2 multiplied by 10-3μg。
Sixthly, sample determination
The results of the measurements performed on 3 batches of samples are shown in Table 4.
TABLE 4 results of content measurement of samples
| Sample batch number | Content of morroniside (mg/g) | Loganin content (mg/g) |
| 181119 | 0.169 | 0.512 |
| 181203 | 0.183 | 0.535 |
| 181218 | 0.188 | 0.527 |
Seventh, investigation of method
The content determination method under the item of dogwood selects acetonitrile-0.3 percent phosphoric acid gradient elution as a mobile phase. Scanning in the wavelength range of 200-400 nm, and selecting 240nm with maximum absorption and less interference peaks to measure morroniside and loganin.
In the research of the sample pretreatment method, the sample is divided into a methanol heating reflux group, a methanol ultrasonic group, a 70% methanol heating reflux group, a 70% methanol ultrasonic group, an ethanol heating reflux group, an ethanol ultrasonic group, a 70% ethanol heating reflux group and a 70% ethanol ultrasonic group, and the content of the component to be detected is inspected. As a result, the content of each component to be detected extracted by the methanol is larger than that of the ethanol; compared with the ultrasonic treatment, the heating reflux extraction has no more chromatographic peak information, and the content difference of the components to be detected is not large, so the ultrasonic treatment by methanol is selected. Meanwhile, the content difference of the components to be detected under different extraction times (30 minutes, 45 minutes and 60 minutes) is considered. As a result, the contents of the 3 groups of samples have no obvious difference, so that the ultrasonic treatment is selected for 30 minutes.
The average measurement of morroniside in 3 samples was 0.180mg/g, and the average measurement of loganin was 0.525 mg/g. Considering that the total amount of morroniside and loganin in the pill for tonifying kidney and strengthening bone is not lower than 0.56mg/g based on 80% of average measurement value.
(4) According to the qualitative and quantitative conformity of the traditional Chinese medicine components, the method is used as the basis for judging the qualification of the kidney-tonifying and bone-strengthening pill.
In the patent medicine, the rehmannia root, the prepared rhizome of rehmannia, the teasel root, the red paeony root and the clematis root are added into the medicine by water decoction, the identification and research of the medicines in the patent medicine are tried by adopting thin-layer chromatography, and the thin-layer chromatography effects of aqueous solution, methanol extracting solution, n-butyl alcohol extracting solution, ethyl acetate extracting solution, petroleum ether extracting solution and the like are respectively considered. As a result, paeoniflorin was detected only from the methanol extract of radix Paeoniae Rubra, and it was difficult to detect the specific components from other herbs. The complexity of the ingredients of the traditional Chinese medicine determines that the effective ingredients and contents in the traditional Chinese medicine cannot be completely judged. However, it is sufficient to determine the contents of the ingredients and main substances of the kidney-tonifying and bone-strengthening pill within the disclosure of this example.
The present invention is not limited to the above-mentioned embodiments, and based on the technical solutions disclosed in the present invention, those skilled in the art can make some substitutions and modifications to some technical features without creative efforts according to the disclosed technical contents, and these substitutions and modifications are all within the protection scope of the present invention.
Claims (6)
1. A detection method of traditional Chinese medicine components in a kidney-tonifying and bone-strengthening pill is characterized by comprising the following steps:
(1) grinding the tested patent medicine into powder, observing under a microscope, and primarily judging whether the tested patent medicine contains dogwood, eucommia bark, medlar, malaytea scurfpea fruit, angelica, astragalus, white paeony root, earthworm and amomum fruit;
(2) identifying a thin layer;
(2.1) taking the powder of the tested finished medicine, adding methanol, heating and refluxing, concentrating, then adding the concentrated solution on a neutral alumina column, eluting with methanol, removing the solution and evaporating to dryness, dissolving the residue in water, extracting with water-saturated n-butyl alcohol, evaporating to dryness, adding methanol for dissolution, and preparing a test solution; preparing control medicinal materials of dogwood, astragalus, white paeony root and red paeony root into a control medicinal material solution by the same method in the section; adding methanol into ursolic acid, astragaloside IV and penoniflorin as control solutions; sucking the test solution, the reference solution and the reference solution, spotting on the same silica gel G thin layer plate, performing chromatographic analysis, and observing the consistency of the characteristic spots of the chromatogram of the test solution and the chromatograms of the reference solution and the reference solution;
(2.2) taking the powder of the to-be-tested patent medicine, adding methanol, carrying out ultrasonic treatment and filtering, and concentrating the filtrate to be used as a test solution; preparing radix Angelicae sinensis and fructus Psoraleae in the same way to obtain control medicinal solution; taking ligustilide, psoralen and isopsoralen, and adding methanol respectively to obtain reference solutions; sucking the test solution, the reference medicinal material solution and the reference solution, spotting on the same silica gel G thin-layer plate, performing fluorescence chromatography, and observing the consistency of the characteristic spots of the chromatogram of the test solution, the chromatogram of the reference medicinal material solution and the chromatogram of the reference solution;
(2.3) taking the powder of the tested patent medicine, adding water, heating and refluxing, filtering and concentrating, shaking and extracting by using ethyl acetate, and concentrating again to obtain a test solution I; taking the powder of the tested patent medicine, adding ethyl acetate, filtering after ultrasonic treatment, and concentrating the filtrate to be used as a test solution II; preparing a fructus lycii reference medicinal material into a reference medicinal material solution according to the preparation method of the test solution I; preparing a reference medicinal material solution from eucommia ulmoides according to the preparation method of the test solution II; sucking a test solution I, a test solution II and a reference medicinal material solution, respectively dropping the test solution I, the test solution II and the reference medicinal material solution on the same silica gel G thin-layer plate, performing fluorescence chromatographic analysis by taking toluene-ethyl acetate-formic acid as a developing agent, and observing the consistency of characteristic spots of the chromatogram of the test solution and the chromatogram of the reference medicinal material solution;
(3) quantitative analysis of morroniside and loganin: performing gradient elution with C18 chromatographic column with acetonitrile as mobile phase A and phosphoric acid solution with mass content of 0.3% as mobile phase B, and performing HPLC analysis at 240nm wavelength; acquiring peak areas of morroniside and loganin, and calculating the contents of morroniside and loganin according to the following regression equation:
morroniside Y =16194X +4280 regression coefficient 0.9997
Loganin Y =16186X +9019 regression coefficient 1
Wherein, Y is peak area, X is content, unit mug/mL;
calculating the sum of the morroniside and loganin content according to the value of X, and determining that the total amount of morroniside and loganin in the patent medicine is not less than 0.56 mg/g;
(4) according to the qualitative and quantitative conformity of the traditional Chinese medicine components, the method is used as the basis for judging the qualification of the kidney-tonifying and bone-strengthening pill.
2. The method for detecting the traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill according to claim 1, which is characterized in that: in the step (1), when the observation is carried out under a microscope,
the surface of the epidermal cell of the pericarp is polygonal or rectangular, the diameter is 16-30 mu m, the vertical peripheral wall is thickened in a bead shape, the outer flat peripheral wall is thickened in a granular cutin shape, the cell cavity contains light orange yellow substances, the mesocarp cell is orange brown and is more shrunken, and the pericarp cell is primarily judged to contain the dogwood;
the rubber threads are stripped or twisted into balls, the surface of the rubber threads is granular, and the rubber threads are primarily judged to contain eucommia;
the surface of the pericarp stone cell is irregular and polygonal, the wall thickness is thick, the wavelike bending is realized, the striations are clear, and the pericarp stone cell is primarily judged to contain the medlar;
the fibers are bundled or scattered, the diameter is 8-30 mu m, the wall thickness is thick, longitudinal cracks are formed on the surface, the primary wall and the secondary wall are separated, the two ends of the primary wall and the secondary wall are always broken into whisker shapes or flat sections, and the astragalus membranaceus is preliminarily judged to be contained;
the phloem parenchyma cells are spindle-shaped, the wall is slightly thick, the surface has extremely fine obliquely staggered textures, the transverse septa of the phloem parenchyma cells can be seen sometimes, and the Chinese angelica is preliminarily judged;
calcium oxalate clusters with the diameter of 11-35 mu m exist in parenchymal cells and are arranged in rows, or one cell contains a plurality of clusters, and the cells are preliminarily judged to contain white paeony roots;
longitudinal furrows are observed on the side surface of the gridlike cells, 1 bright band is positioned at the edge close to the upper side, the top surface is polygonal, the cell cavity is extremely small, the pore furrows are fine, and the gridlike cells are preliminarily judged to contain the fructus psoraleae;
the endothelial sclereid cells are reddish brown or yellowish brown, the surfaces of the endothelial sclereid cells are polygonal, the wall thicknesses of the endothelial sclereid cells are not lignified, the intracellular cavities contain siliceous blocks, the sections of the endothelial sclereid cells are 1 row of grid cells, the inner walls and the side walls of the endothelial sclereid cells are extremely thick, the intracellular cavities are arranged on the outer sides, and the endothelial sclereid blocks are preliminarily judged to contain fructus amomi;
the epidermal cells are brownish yellow, the cell boundary is not obvious, dark brown pigment particles are distributed, and the earthworm is initially judged to be contained.
3. The method for detecting traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill according to claim 1, which is characterized in that the specific method in the step (2.1) is as follows:
taking 5g of test sample powder, adding 50mL of methanol, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 15mL, adding the filtrate onto a 5g neutral alumina column with a 100-120 mesh sieve, eluting with 100mL of 40% methanol aqueous solution, collecting the eluent, evaporating to dryness, dissolving the residue in 30mL of water, extracting for 2 times with water-saturated n-butanol by shaking for 20mL each time, combining the n-butanol solution, evaporating to dryness, and dissolving the residue in 1mL of methanol to obtain a test sample solution; preparing control medicinal materials of Corni fructus, radix astragali, radix Paeoniae alba, and radix Paeoniae Rubra each 1g, and making into control medicinal solution respectively by the same method; adding methanol into ursolic acid, astragaloside IV and penoniflorin as reference substances to obtain 1mg solution per 1mL as reference substance solution; in the thin layer chromatography test, sucking 5 μ L of the sample solution, 2 μ L of the reference solution and 2 μ L of the reference solution, respectively dropping on the same silica gel G thin layer plate, developing with the lower layer solution of dichloromethane-methanol-water solution at volume ratio of 13: 6: 2 as developing agent, taking out, air drying, spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and observing the consistency of the characteristic spots of the chromatogram of the sample solution and the chromatogram of the reference solution and the reference solution.
4. The method for detecting traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill according to claim 1, which is characterized in that the specific method in the step (2.2) is as follows:
taking 2g of test sample powder, adding 30mL of methanol, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to be used as a test sample solution; preparing 1g of radix Angelicae sinensis and fructus Psoraleae control medicinal materials respectively, and making into control medicinal solution respectively; then taking ligustilide, psoralen and isopsoralen as reference substances, and adding methanol to obtain 1mg solution per 1mL as reference substance solution; when performing thin layer chromatography, sucking 2 μ L of each of the six solutions, respectively dropping on the same silica gel G thin layer plate, developing with n-hexane-ethyl acetate solution at volume ratio of 7: 2 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color.
5. The method for detecting traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill according to claim 1, which is characterized in that the specific method in the step (2.3) is as follows:
taking 5g of test sample powder, adding 50mL of water, heating and refluxing for 1 hour, filtering, concentrating the filtrate to 20mL, extracting with ethyl acetate by shaking for 2 times, 15mL each time, combining ethyl acetate solutions, and concentrating to 1mL to obtain a test sample solution I; taking 2g of test sample powder, adding 30mL of ethyl acetate, carrying out ultrasonic treatment for 30 minutes, filtering, and concentrating the filtrate to 1mL to obtain a test sample solution II; taking 1g of fructus Lycii reference material, and making into reference material solution according to the preparation method of the test solution I; taking 1g of eucommia ulmoides as a reference medicinal material, and preparing a reference medicinal material solution according to a preparation method of a test solution II; when performing thin layer chromatography, sucking 5 μ L of sample solution I, 5 μ L of sample solution II and 2 μ L of control solution, respectively dropping on the same silica gel G thin layer plate, developing with toluene-ethyl acetate-formic acid solution at volume ratio of 15: 8: 1.5 as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp; and judging whether the positions of the chromatogram of the test sample, the reference medicinal material and the reference substance are corresponding to the fluorescent spots with the same color.
6. The method for detecting traditional Chinese medicine components in the kidney-tonifying and bone-strengthening pill according to any one of claims 1 to 5, wherein the quantitative analysis of morroniside and loganin in the step (3) is specifically performed by:
taking 5g of test sample powder, precisely weighing, placing in a conical flask with a plug, precisely adding 50mL of methanol, and weighing; ultrasonic processing with power of 300W and frequency of 50kHz for 30 minutes, cooling, weighing, supplementing the weight loss with methanol, shaking up, and filtering; respectively and precisely sucking 10 mu L of test solution, injecting into a liquid chromatograph, and carrying out chromatography with Agilent Zorbax SB-C18 chromatographic column; taking acetonitrile as a mobile phase A, taking a phosphoric acid solution with the weight content of 0.3% as a mobile phase B, and carrying out gradient elution; the flow rate was 1.0mL per minute; under the column temperature of 30 ℃, the detection wavelength is 240nm, the peak areas of the morroniside and the loganin are obtained, and then calculation is carried out.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111458447A (en) * | 2020-05-11 | 2020-07-28 | 贵州中医药大学 | Chinese wolfberry quality detection method based on spectral efficiency relationship |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1879756A (en) * | 2006-04-30 | 2006-12-20 | 石药集团中奇制药技术（石家庄）有限公司 | Quality control method of kidney-replenishing blood-nourishing soft capsule |
| CN1903345A (en) * | 2006-08-10 | 2007-01-31 | 李国柱 | Traditional Chinese medicine pills for tonifying kidney and strengthening bones for treating osteoarthritis and prodn. method thereof |
| CN101574477A (en) * | 2009-05-18 | 2009-11-11 | 唐弟光 | Method for controlling quality of menstruation regulation and macula removal tablet |
| CN101822765A (en) * | 2010-01-28 | 2010-09-08 | 江西新赣江药业有限公司 | Quality inspection method of traditional Chinese medicine compound preparation with functions of tonifying kidney, strengthening bones, enriching blood and being beneficial to sperm |
| CN102233097A (en) * | 2010-05-07 | 2011-11-09 | 郑州福瑞堂制药有限公司 | Quality control method for Zengguang capsules |
| CN102520103A (en) * | 2011-12-19 | 2012-06-27 | 辽宁省食品药品检验所 | Contrast Chinese patent medicine extract of pills of six ingredients with rehmannia, and preparation method and application thereof |
| CN102879516A (en) * | 2012-09-25 | 2013-01-16 | 南京中医药大学 | Method for identifying Buyang Huanwu soup and measuring content of Buyang Huanwu soup |
| CN106680399A (en) * | 2017-01-20 | 2017-05-17 | 天津市第中心医院 | Method for simultaneously determining content of morroniside, loganin, cor-nuside, oleanolic acid and ursolic acid |
-
2019
- 2019-12-30 CN CN201911395931.5A patent/CN111024877A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1879756A (en) * | 2006-04-30 | 2006-12-20 | 石药集团中奇制药技术（石家庄）有限公司 | Quality control method of kidney-replenishing blood-nourishing soft capsule |
| CN1903345A (en) * | 2006-08-10 | 2007-01-31 | 李国柱 | Traditional Chinese medicine pills for tonifying kidney and strengthening bones for treating osteoarthritis and prodn. method thereof |
| CN101574477A (en) * | 2009-05-18 | 2009-11-11 | 唐弟光 | Method for controlling quality of menstruation regulation and macula removal tablet |
| CN101822765A (en) * | 2010-01-28 | 2010-09-08 | 江西新赣江药业有限公司 | Quality inspection method of traditional Chinese medicine compound preparation with functions of tonifying kidney, strengthening bones, enriching blood and being beneficial to sperm |
| CN102233097A (en) * | 2010-05-07 | 2011-11-09 | 郑州福瑞堂制药有限公司 | Quality control method for Zengguang capsules |
| CN102520103A (en) * | 2011-12-19 | 2012-06-27 | 辽宁省食品药品检验所 | Contrast Chinese patent medicine extract of pills of six ingredients with rehmannia, and preparation method and application thereof |
| CN102879516A (en) * | 2012-09-25 | 2013-01-16 | 南京中医药大学 | Method for identifying Buyang Huanwu soup and measuring content of Buyang Huanwu soup |
| CN106680399A (en) * | 2017-01-20 | 2017-05-17 | 天津市第中心医院 | Method for simultaneously determining content of morroniside, loganin, cor-nuside, oleanolic acid and ursolic acid |
Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会: "《中华人民共和国药典 2015年版 一部》", 30 June 2015, 中国医药科技出版社 * |
| 姜大成: "《中药鉴定学》", 31 January 2016, 中国农业大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111458447A (en) * | 2020-05-11 | 2020-07-28 | 贵州中医药大学 | Chinese wolfberry quality detection method based on spectral efficiency relationship |
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