CN110927281B - Detection method of pharmaceutical composition Rupishu capsule for treating hyperplasia of mammary glands and mastitis - Google Patents
Detection method of pharmaceutical composition Rupishu capsule for treating hyperplasia of mammary glands and mastitis Download PDFInfo
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Abstract
The invention relates to a detection method of a pharmaceutical composition for treating hyperplasia of mammary glands and mastitis, in particular to a detection method of a Rupishu capsule for treating hyperplasia of mammary glands and mastitis. The detection method is sensitive, has strong specificity, is accurate and has good reproducibility, can make up the defects of the existing method, effectively controls the quality of the medicine, and associates the quality of the medicine with the effective components of clinical treatment, thereby achieving the standards of stability, controllability, high efficiency and safety and further better meeting the medical needs.
Description
Technical Field
The invention belongs to the technical field of pharmacy, in particular relates to a detection method of a pharmaceutical composition for treating hyperplasia of mammary glands and mastitis, and particularly relates to a detection method of a Rupishu capsule for treating hyperplasia of mammary glands and mastitis.
Background
The mammary gland hyperplasia and mastitis are the most common breast diseases of women, the incidence rate of the diseases is on the trend of increasing year by year in recent years, and the ages are also getting lower and lower; epidemiological investigations have shown that about 70% of adult women between 20 and 55 years of age have different degrees of mammary hyperplasia. Among all breast diseases, hyperplasia of mammary glands is the most common, accounting for more than 90% of breast outpatients, and the investigation of asymptomatic women in the childbearing age indicates that the number of patients with various breast diseases reaches 52.4%, wherein the number of women suffering from hyperplasia of mammary glands is as high as 49.7%. The incidence of hyperplasia of mammary glands is much higher than that of other chronic common diseases of women, and hyperplasia of mammary glands accounts for approximately 75 percent of all mammary glands. At present, more than 1000 million cases of new breast hyperplasia are suffered in China every year, and the onset age is in a youthful trend. The Rupishu capsule is produced by Xian Qian He pharmaceutical industry GmbH, the prescription is composed of 7 medicines of salvia miltiorrhiza, snakegourd fruit, red paeony root, rhizoma bolbostemmae, dandelion, corydalis tuber and radix bupleuri, and the capsule is clinically used for treating hyperplasia of mammary glands and mastitis caused by stagnation of liver qi and mutual resistance of toxin and blood stasis. The efficacy of the medicine is seriously trusted and favored by patients and medical workers for more than ten years.
However, in the existing detection method of the Rupishu capsule, only thin-layer identification is carried out on paniculate Bolbostemma rhizome, adjunctive drug Salvia miltiorrhiza and radix Paeoniae Rubra, and content determination of paeoniflorin is carried out on adjunctive drug radix Paeoniae Rubra; the content of monarch drugs and ministerial drugs in the prescription is not measured, the specificity of the existing quality control standard is poor, the quality of the drugs cannot be comprehensively reflected, and the clinical curative effect index cannot be controlled, so that a detection method of the product is necessary to be further perfected, and the control index is associated with the clinical curative effect, so that the quality and the curative effect of the product are further ensured.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, and provides a detection method of a pharmaceutical composition Rupishu capsule for treating hyperplasia of mammary glands and mastitis, wherein the content of quercetin which is an active ingredient in monarch drug pericarpium trichosanthis and ministerial drug dandelion is measured.
In order to achieve the purpose, the invention adopts the following technical scheme:
a detection method of a pharmaceutical composition Rupishu capsule for treating hyperplasia of mammary glands and mastitis comprises the following seven medicinal materials: saviae Miltiorrhizae radix, pericarpium Trichosanthis, radix Paeoniae Rubra, paniculate Bolbostemma rhizome, herba Taraxaci, rhizoma corydalis, and bupleuri radix; the method for measuring the content of quercetin by adopting liquid chromatography comprises the following steps:
(1) chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; gradient elution is carried out by taking an organic solvent as phase A and a dilute acid aqueous solution with a certain concentration as phase B, wherein the organic solvent is selected from methanol, acetonitrile or acetonitrile-methanol mixed solution, and the dilute acid aqueous solution is selected from phosphoric acid aqueous solution with weight volume percentage concentration of 0.01-1.0% or acetic acid aqueous solution with weight volume percentage concentration of 0.01-0.5%; the column temperature is 20-40 ℃; an ultraviolet detector with the detection wavelength of 360-380 nm; the flow rate is 0.8ml/min-1.2 ml/min; the column temperature is 20-40 ℃; the number of theoretical plates is not less than 3000 calculated by quercetin;
preferably, the phase A is methanol, and the phase B is phosphoric acid aqueous solution with the volume percentage concentration of 0.2%; the flow rate is 1.0 ml/min; the column temperature is 30 ℃; the detection wavelength is 370 nm. Preferably, when the gradient elution is performed, the gradient elution procedure is as follows: 0-10 min, 50% A; 10-15 min, 50-65% A; 15-25 min, 65% A; 25-28 min, 50% A.
Gradient elution was performed as specified in the table below;
| time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0～10 | 50 | 50 |
| 10～15 | 50→65 | 50→35 |
| 15～25 | 65 | 35 |
| 25～28 | 50 | 50 |
(2) Preparation of control solutions
Taking a quercetin reference substance, precisely weighing, and adding methanol to prepare a solution containing 30 μ g quercetin per lml;
(3) preparation of test solution
Taking the content of the capsule for treating hyperplasia of mammary glands, grinding, weighing 1.0-3.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 25-100ml of water, weighing, heating and refluxing for 0.5-1.5 hours, cooling, weighing again, supplementing the weight loss with water, shaking up, and filtering; precisely measuring 5-15ml of subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, each time 5-15ml, combining ethyl acetate solutions, recovering solvent under reduced pressure to dry, adding mobile phase into residue to dissolve, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting subsequent filtrate;
preferably, the content of the capsule for treating hyperplasia of mammary glands is taken, ground, weighed to be 2.0g, precisely weighed, placed in a conical flask with a plug, precisely added with 50ml of water, weighed, heated and refluxed for 1 hour, cooled, weighed again, supplemented with water to reduce the weight, shaken evenly and filtered; precisely measuring 10ml of the subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, each time 10ml, mixing ethyl acetate solutions, recovering solvent under reduced pressure to dryness, adding mobile phase into residue to dissolve, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting the subsequent filtrate;
(4) measurement of
Precisely sucking 10-20 μ l of each of the reference solution and the sample solution, respectively, injecting into high performance liquid chromatograph, and measuring to obtain measurement result.
The prescription of the capsule for treating mammary nodules in the invention is as follows: 450g of pericarpium trichosanthis (monarch drug), 150g of rhizoma bolbostemmae (ministerial drug), 450g of dandelion (ministerial drug), 225g of salvia miltiorrhiza (adjuvant drug), 225g of red paeony root (adjuvant drug), 135g of rhizoma corydalis (adjuvant drug) and 135g of radix bupleuri (conductant drug), the formula takes the pericarpium trichosanthis as the monarch drug, which is the essential drug for treating acute mastitis and nodules of breast, and can promote qi circulation, relieve depression, soften hardness and dissipate stagnation; paniculate Bolbostemma rhizome is taken as a ministerial drug, which is good at treating acute mastitis and nodules of breast, and the herbal medicine is said from the new records: tubei fritillary bulb for treating surgical phlegm toxin. Shanxi Chinese herbal medicine cloud: it is good at treating acute mastitis initial stage and carbuncle swelling, and has strong effects of removing toxic substance and resolving hard mass, eliminating carbuncle swelling, and assisting monarch drug to strengthen its effects of softening hard mass and resolving hard mass; the dandelion is also taken as a ministerial drug, has the effects of clearing heat and removing toxicity, and reducing swelling and dissipating stagnation, and is a key drug for treating acute mastitis and a common drug for treating nodules of breast. The snakegourd peel has the effects of clearing heat and eliminating phlegm, and benefiting qi and relieving chest stuffiness, and is mainly used for symptoms such as phlegm-heat cough, chest distress and hypochondriac pain. Modern researches show that the pericarpium trichosanthis mainly contains bioactive components such as organic acid, grease, flavone and glycosides thereof, saccharides, alkaloid, amino acid, sterol and glycosides thereof and the like, and has various effects of dilating coronary artery, resisting arrhythmia, resisting oxidative stress, inhibiting platelet aggregation, reducing cholesterol, resisting bacteria, diminishing inflammation, resisting cancer and the like. The dandelion is bitter and sweet in taste and cold in nature, has the effects of clearing heat and removing toxicity, inducing diuresis and resolving masses, is used for treating various symptoms of sore, carbuncle and ulcer, red swelling and heat toxin and is particularly quick in effect for treating women's human breast cancer and swollen breast toxin. Modern pharmacological research shows that the dandelion has the functions of bacteriostasis, anti-inflammation, antioxidation, anti-tumor, blood sugar reduction and blood fat reduction. Mainly contains chemical components such as triterpenes, flavonoids, coumarins, sesquiterpene lactones and the like. The flavonoid compounds of snakegourd peel and dandelion have higher quercetin content. Quercetin is one of effective components of pericarpium Trichosanthis and herba Taraxaci, has various biological activities and high medicinal value, has effects of resisting oxidation and scavenging free radicals, and also has anticancer, antiinflammatory, antibacterial, antivirus, blood sugar and blood pressure lowering, immunity regulating and cardiovascular protecting effects etc.
Compared with the prior art, the invention has the following beneficial technical effects:
according to the method for measuring the content of the Rupishu capsule, only thin-layer identification is carried out on the ministerial drugs of rhizoma bolbostemmae, the adjunctive drugs of radix salviae miltiorrhizae and radix paeoniae rubra in the original quality standard, and the content measurement of paeoniflorin is carried out on the adjunctive drugs of radix paeoniae rubra; the content of monarch drugs and ministerial drugs in the prescription is not measured, the specificity of the existing quality control standard is poor, and the quality and the curative effect of the product cannot be ensured. The detection method is further perfected on the basis, the content of the quercetin which is the active ingredient in the monarch drug snakegourd peel and the ministerial drug taraxacum is measured, the detection method is sensitive, high in specificity, accurate and good in reproducibility, the defects of the existing method can be overcome, the quality of the drugs can be effectively controlled, the stable, controllable, efficient and safe standards can be achieved, and the basis is provided for improving the quality standard of the variety.
Drawings
The invention is further described with reference to the accompanying drawings in which:
FIG. 1 is a blank solution chromatogram in a specificity test under chromatographic conditions according to the present invention;
FIG. 2 is a chromatogram of a control solution in a specificity test under chromatographic conditions according to the present invention;
FIG. 3 is a chromatogram of a negative sample solution in a specificity test under chromatographic conditions according to the present invention;
FIG. 4 is a chromatogram of a sample solution in a specificity test under chromatographic conditions according to the present invention;
FIG. 5 is a first pin chromatogram of instrument precision under chromatographic conditions in accordance with the present invention;
FIG. 6 is a second chromatogram of the instrument under chromatographic conditions;
FIG. 7 is a third chromatogram of the instrument precision under chromatographic conditions;
FIG. 8 is a fourth chromatogram of the instrument precision under chromatographic conditions;
FIG. 9 is a fifth chromatogram of the instrument under chromatographic conditions;
FIG. 10 is a sixth chromatogram for instrument precision under chromatographic conditions in accordance with the present invention;
FIG. 11 is a linear test pattern of the present invention at a concentration of 17.22. mu.g/ml under chromatographic conditions;
FIG. 12 is a linear test pattern of the present invention at a concentration of 34.44. mu.g/ml under chromatographic conditions;
FIG. 13 is a linear test pattern of 68.88 μ g/ml concentration under chromatographic conditions in accordance with the present invention;
FIG. 14 is a linear test pattern of 86.10 μ g/ml concentration under chromatographic conditions in accordance with the present invention;
FIG. 15 is a linear test pattern of concentration 103.32 μ g/ml under chromatographic conditions according to the invention'
FIG. 16 is a linear relationship under chromatographic conditions for the present invention.
Detailed Description
The invention is illustrated below with reference to specific examples. It will be understood by those skilled in the art that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention in any way. The experimental procedures in the following examples are conventional unless otherwise specified. The raw materials, reagents and materials used in the following examples are all commercially available products unless otherwise specified.
Example 1: prescription of Rupishu capsule and its preparation method
Prescription: 450g of snakegourd peel, 150g of rhizoma bolbostemmae, 450g of dandelion, 225g of salvia miltiorrhiza, 225g of red paeony root, 135g of rhizoma corydalis and 135g of radix bupleuri
The preparation method comprises the following steps: pulverizing rhizoma corydalis into fine powder; extracting the red sage root and the red paeony root twice by adding ethanol, adding 5 times of ethanol for each time, extracting for 2 hours respectively, combining ethanol extracting solutions, filtering, recovering the ethanol from the filtrate, and concentrating to form thick paste with the relative density of 1.33-1.36 (60 ℃); decocting the rest four materials such as the pericarpium trichosanthis and the like in water for three times, adding 10 times of water for the first time, decocting for 2 hours, adding 8 times of water for the second time, decocting for 1.5 hours, adding 6 times of water for the third time, decocting for 1 hour, merging decoction, filtering, concentrating filtrate under reduced pressure to obtain thick paste with the relative density of 1.33-1.36 (60 ℃), merging the thick paste, adding the fine powder of rhizoma corydalis, 12g of silicon dioxide and 8g of pregelatinized starch, stirring uniformly, drying in vacuum, crushing into fine powder, and filling into capsules to prepare 1000 capsules.
Example 2: selection of wavelength and mobile phase for measuring quercetin content in Rupishu capsule
2.1 selection of wavelength
The quercetin control solution is subjected to wavelength scanning within the range of 200-500nm, the result shows that the maximum absorption exists at 254nm and 370nm, and the 370nm with less interference is selected as the detection wavelength considering that the absorbance of other components in the prescription at 254nm is also larger and the interference is more.
2.2 selection of the Mobile phase
The preliminary tests showed that the tests were carried out using different gradients of acetonitrile-water-1.0% phosphoric acid aqueous solution, methanol-4% phosphoric acid aqueous solution, methanol-acetonitrile-0.2% phosphoric acid aqueous solution, acetonitrile-0.2% HAc aqueous solution, methanol-0.2% HAc aqueous solution, and methanol-water. The results show that the separation effect is best with a linear gradient of methanol-0.1% phosphoric acid in water, with the baseline being the most stable. The methanol-0.1% phosphoric acid aqueous solution mobile phase system is further optimized.
In order to effectively separate quercetin in a test product and meet the requirements of peak shape, theoretical plate number and the like, mobile phases with different proportions are experimentally investigated, and finally the fluidity described in the invention content is selected, which is specifically shown in the following table 1.
Table 1 mobile phase selection
Example 3: research on specialization
3.1 preparation of negative samples:
prescription: 150g of rhizoma bolbostemmae, 225g of salvia miltiorrhiza, 225g of red paeony root, 135g of rhizoma corydalis and 135g of radix bupleuri
The preparation method comprises the following steps: pulverizing rhizoma corydalis into fine powder; extracting the red sage root and the red paeony root twice by adding ethanol, adding 5 times of ethanol for each time, extracting for 2 hours respectively, combining ethanol extracting solutions, filtering, recovering the ethanol from the filtrate, and concentrating to form thick paste with the relative density of 1.33-1.36 (60 ℃); decocting the other two medicines such as the paniculate Bolbostemma rhizome and the like in water for three times, adding 10 times of water for the first time, decocting for 2 hours, adding 8 times of water for the second time, decocting for 1.5 hours, adding 6 times of water for the third time, decocting for 1 hour, mixing decoctions, filtering, concentrating the filtrate under reduced pressure to obtain thick paste with the relative density of 1.33-1.36 (60 ℃), mixing the thick pastes, adding the fine powder of corydalis tuber, 12g of silicon dioxide and 8g of pregelatinized starch, stirring uniformly, drying in vacuum, and crushing into fine powder to obtain the traditional Chinese medicine.
3.2 preparation of negative sample solution:
weighing 2.0g of negative sample, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of water, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the weight loss with water, shaking up, and filtering. Precisely measuring 10ml of the subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, 10ml each time, mixing ethyl acetate solutions, recovering solvent under reduced pressure, dissolving the residue with mobile phase, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting the subsequent filtrate.
3.3 preparation of test solution:
taking the content of the Rupishu capsule, grinding, weighing 2.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of water, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the lost weight with water, shaking up, and filtering. Precisely measuring 10ml of the subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, 10ml each time, mixing ethyl acetate solutions, recovering solvent under reduced pressure, dissolving the residue with mobile phase, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting the subsequent filtrate.
3.4 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
preparation of control solutions:
taking a quercetin reference substance, precisely weighing, and adding methanol to prepare a solution containing 30 μ g quercetin per lml;
preparation of a blank solution: methanol was taken directly as a blank solution.
And (3) determination:
precisely sucking 10 μ l of each of the negative sample solution, the reference solution, the sample solution and the blank solution, injecting into a high performance liquid chromatograph, and measuring.
The results show that: in the chromatogram of the test solution, the same chromatographic peak is arranged at the corresponding position of the chromatographic peak of the quercetin reference substance; negative and blank reagents are not interfered, which shows that the chromatographic condition has good specificity. See in particular figures 1-4 below.
Example 4: repeatability test
4.1 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
4.2 precisely absorbing 10 mul of the same sample solution, injecting into a liquid chromatograph, repeatedly injecting for 6 times, and calculating the peak area RSD.
4.3 the results show that the RSD value of the peak area is 0.3%, less than 2.0%; indicating good precision. The results are shown in tables 2-3 and FIGS. 5-10 below.
TABLE 2 precision test data sheet
TABLE 3 detailed data table of precision test
Example 5: linearity, range
5.1 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
5.2 the control solutions were aspirated as: 17.22. mu.g/ml, 34.44. mu.g/ml, 68.88. mu.g/ml, 86.10. mu.g/ml, 103.32. mu.g/ml each 10. mu.l were injected into a liquid chromatograph, a chromatogram was recorded, a linear regression analysis was performed on the concentration (x) by the peak area (y), a linear relationship graph was drawn, and a linear equation was reported. The results are shown in Table 4 and FIGS. 11-16.
TABLE 4 Linear relationship test data sheet
5.3 results show that under the condition of the sample amount of 10 mul, the quercetin has a good linear relation with the peak area within the concentration range of 17.22-103.32 mug/ml.
Example 6: accuracy (recovery)
6.1 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
6.2 preparation of control solutions: precisely weighing quercetin control, and adding methanol to obtain control solution with concentration of 34.44 μ g/ml;
6.3 preparation of test solution: taking the content of the Rupishu capsule, grinding, weighing 1.0g and 9 parts respectively, precisely weighing, placing in a conical flask with a plug, precisely weighing 9 parts of quercetin reference substances respectively, sequentially adding into the conical flask after sample addition, precisely adding 50ml of water respectively, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the weight loss with water, shaking uniformly, and filtering. Precisely measuring 10ml of the subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, 10ml each time, mixing ethyl acetate solutions, recovering solvent under reduced pressure, dissolving the residue with mobile phase, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting the subsequent filtrate.
Percent recovery is (C-A)/BX 100%
Wherein A is the measured component content of the test sample; b is the amount of the added reference substance; c is an observed value. The results are shown in Table 5.
TABLE 5 recovery test data sheet
The standard requires that: the recovery rate is 85-110%, and the RSD is less than 4.0%; the experimental result shows that the method has good accuracy.
Example 7: intermediate precision
7.1 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
7.2 in the same laboratory, different experimenters A and B perform experiments by using different chromatographs on different dates, recording chromatograms and calculating the content. The results are shown in Table 6.
TABLE 6 precision test data sheet
7.3 the result shows that the invented patented method is good in precision.
Example 8: durability
8.1 chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; the chromatographic conditions described in example 2 and in Table 1 No. 4 are used, the theoretical plate number should not be less than 3000 calculated as quercetin;
8.2 taking the same sample solution, standing at room temperature for 0, 4, 8, 12, 24 and 36 hours respectively, and precisely sucking 10 mul to inject into a high performance liquid chromatograph; the results are shown in Table 7.
Table 7 stability test data table
8.3, the result shows that the RSD value of the sample solution is 0.9 percent within 24 hours, the RSD value of the peak area is 1.9 percent, and the RSD values are all less than 2.0 percent; the test solution was stable for 36h at room temperature.
Example 9: content determination and stability test of quercetin in Rupishu capsule
Samples of the Rupishu capsule mass production (lot numbers: 20180401, 20180402, 20180403, manufactured by Seikagaku pharmaceutical industries, Ltd.) were subjected to accelerated tests and room temperature standing tests. Accelerated experiment, placing the sample in a closed environment with relative humidity of 75% +/-5% and temperature of 40 +/-2 ℃ for 6 months, sampling in 0, 1, 2, 3 and 6 months for character observation and microorganism limit detection; room temperature standing experiment: the sample was placed in a closed environment with a relative humidity of 60% +/-5% and a temperature of 25 deg.C + -2 deg.C for 3 months, and sampled and examined at 0, 3, 6, 12, and 18 months, and the test results are shown in Table 8.
TABLE 8 stability test data sheet for quercetin content in Rupishu capsule
The experimental result shows that the content of the quercetin does not change significantly after the test is accelerated for 6 months and the test is placed at room temperature for 18 months, meets the requirement, and shows that the method for detecting the content of the quercetin is applied to more effectively control the quality of the medicine and is considered to be included in the quality standard in the next step.
In summary, the following steps: the method for measuring the content of the quercetin in the Rupishu capsule determines the scientificity of the method from the aspects of a repetitive chromatographic expression chart, a test sample solution stability chart, instrument precision, recovery rate, linearity, specificity and the like. The content determination of monarch drugs and ministerial drugs of the preparation is increased, the safety and effectiveness of the preparation are ensured, the defects of the prior art are overcome, and the detection accuracy is further improved.
The embodiments given above are preferable examples for implementing the present invention, and the present invention is not limited to the above-described embodiments. Any non-essential addition and replacement made by the technical characteristics of the technical scheme of the invention by a person skilled in the art belong to the protection scope of the invention.
Claims (2)
1. A detection method of a pharmaceutical composition Rupishu capsule for treating hyperplasia of mammary glands and mastitis is characterized in that the content of quercetin is determined by adopting a liquid chromatography, and the method specifically comprises the following steps:
(1) chromatographic conditions and System suitability test
Octadecylsilane chemically bonded silica is used as a filling agent; taking methanol as phase A and phosphoric acid aqueous solution with the volume percentage concentration of 0.2% as phase B, and performing gradient elution, wherein the gradient elution procedure is as follows: 0-10 min, 50% A; 10-15 min, 50-65% A; 15-25 min, 65% A; 25-28 min, 50% A; the flow rate is 1.0 ml/min; the column temperature is 30 ℃; the detection wavelength is 370 nm;
(2) preparation of control solutions
Taking a quercetin reference substance, precisely weighing, and adding methanol to prepare a solution containing 30 μ g quercetin per lml;
(3) preparation of test solution
Taking the content of the capsule for treating hyperplasia of mammary glands, grinding, weighing 1.0-3.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 25-100ml of water, weighing, heating and refluxing for 0.5-1.5 hours, cooling, weighing again, supplementing the weight loss with water, shaking up, and filtering; precisely measuring 5-15ml of subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, each time 5-15ml, combining ethyl acetate solutions, recovering solvent under reduced pressure to dry, adding mobile phase into residue to dissolve, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting subsequent filtrate;
(4) measurement of
Precisely sucking 10-20 μ l of each of the reference solution and the sample solution, respectively, injecting into high performance liquid chromatograph, and measuring to obtain measurement result.
2. The method for detecting the pharmaceutical composition Rupishu capsule for treating mammary gland hyperplasia and mastitis according to claim 1, wherein the pharmaceutical composition Rupishu capsule comprises the following components: in the step (3), preparing a test solution: taking the content of the Rupishu capsule, grinding, weighing 2.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of water, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the lost weight with water, shaking up, and filtering; precisely measuring 10ml of the subsequent filtrate, placing in a separating funnel, adding ethyl acetate, extracting for 3 times, 10ml each time, mixing ethyl acetate solutions, recovering solvent under reduced pressure, dissolving the residue with mobile phase, transferring to a 10ml measuring flask, adding mobile phase to scale, shaking, filtering, and collecting the subsequent filtrate.
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| CN110927281B true CN110927281B (en) | 2022-04-19 |
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| CN102641407A (en) * | 2012-04-18 | 2012-08-22 | 皖南医学院 | Preparing method and detecting method for fructus trichosanthis granules |
| CN103033585A (en) * | 2011-09-30 | 2013-04-10 | 西安千禾药业有限责任公司 | Method for detecting medicaments for treating mammitis and hyperplasia of mammary glands |
| KR20180011473A (en) * | 2015-02-11 | 2018-02-01 | 한양대학교 에리카산학협력단 | Method for quality evaluation of hovenia dulcis extract |
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| CN103033585A (en) * | 2011-09-30 | 2013-04-10 | 西安千禾药业有限责任公司 | Method for detecting medicaments for treating mammitis and hyperplasia of mammary glands |
| CN102641407A (en) * | 2012-04-18 | 2012-08-22 | 皖南医学院 | Preparing method and detecting method for fructus trichosanthis granules |
| KR20180011473A (en) * | 2015-02-11 | 2018-02-01 | 한양대학교 에리카산학협력단 | Method for quality evaluation of hovenia dulcis extract |
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| RP-HPLC法测定柴胡中四种黄酮类成分的含量;岳志劲等;《山西大同大学学报(自然科学版)》;20100831;第26卷(第4期);第35-38页 * |
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